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Journal articles on the topic 'Expression plasmid'

Author: Grafiati

Published: 4 June 2021

Last updated: 11 February 2022

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1

Mitchell-Felton, Heather, and Susan C. Kandarian. "Normalization of muscle plasmid uptake by Southern blot: application to SERCA1 promoter analysis." American Journal of Physiology-Cell Physiology 277, no. 6 (1999): C1269—C1276. http://dx.doi.org/10.1152/ajpcell.1999.277.6.c1269.

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Direct injection of plasmid DNA into muscle allows the study of promoters in a physiological environment. Because of the variability of reporter gene activity, attempts have been made to normalize activity to muscle plasmid uptake by coinjection of a second “control” plasmid whose reporter gene is constitutively expressed by a viral promoter. The purpose of this study was to evaluate the use of a control plasmid vs. Southern blot to normalize for differences in uptake of plasmids containing promoter fragments of the skeletal muscle-specific sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA1) gen

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2

Clark, Leann, Isabel Martinez-Argudo, Tom J. Humphrey, and Mark A. Jepson. "GFP plasmid-induced defects in Salmonella invasion depend on plasmid architecture, not protein expression." Microbiology 155, no. 2 (2009): 461–67. http://dx.doi.org/10.1099/mic.0.025700-0.

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We have investigated the impact of plasmids and GFP expression on invasion of cultured epithelial cells by Salmonella enterica Typhimurium strain SL1344. The invasiveness of SL1344 carrying plasmids derived from pBR322, encoding promoterless GFP or constitutively expressed rpsM-GFP, was compared under optimal growth conditions with that of SL1344(pBR322), unmodified SL1344 and a strain with chromosome-integrated rpsM-GFP. The strain carrying pBR322 exhibited normal invasion, but the presence of modified plasmids impaired invasiveness, and impairment was exacerbated by plasmid-encoded chloramph

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3

Galen, James E., Jay Nair, Jin Yuang Wang, et al. "Optimization of Plasmid Maintenance in the Attenuated Live Vector Vaccine Strain Salmonella typhiCVD 908-htrA." Infection and Immunity 67, no. 12 (1999): 6424–33. http://dx.doi.org/10.1128/iai.67.12.6424-6433.1999.

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ABSTRACT The broad objective of the research presented here is to develop a noncatalytic plasmid maintenance system for the stabilization of multicopy expression plasmids encoding foreign antigens in aSalmonella typhi live-vector vaccine strain such as CVD 908-htrA. We have enhanced the maintenance of expression plasmids at two independent levels. First, we removed dependence upon balanced-lethal maintenance systems that involve catalytic enzymes expressed from multicopy plasmids; we accomplished this through incorporation into expression plasmids of a postsegregational killing system based on

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4

Harth, Günter, Saša Masleša-Galić, and Marcus A. Horwitz. "A two-plasmid system for stable, selective-pressure-independent expression of multiple extracellular proteins in mycobacteria." Microbiology 150, no. 7 (2004): 2143–51. http://dx.doi.org/10.1099/mic.0.27113-0.

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Recombinant mycobacteria expressing Mycobacterium tuberculosis extracellular proteins are leading candidates for new vaccines against tuberculosis and other mycobacterial diseases, and important tools both in antimycobacterial drug development and basic research in mycobacterial pathogenesis. Recombinant mycobacteria that stably overexpress and secrete major extracellular proteins of M. tuberculosis in native form on plasmids pSMT3 and pNBV1 were previously constructed by the authors. To enhance the versatility of this plasmid-based approach for mycobacterial protein expression, the Escherichi

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Nakashima, Nobutaka, and Tomohiro Tamura. "Isolation and Characterization of a Rolling-Circle-Type Plasmid from Rhodococcus erythropolis and Application of the Plasmid to Multiple-Recombinant-Protein Expression." Applied and Environmental Microbiology 70, no. 9 (2004): 5557–68. http://dx.doi.org/10.1128/aem.70.9.5557-5568.2004.

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ABSTRACT We isolated, sequenced, and characterized the cryptic plasmid pRE8424 from Rhodococcus erythropolis DSM8424. Plasmid pRE8424 is a 5,987-bp circular plasmid; it carries six open reading frames and also contains cis-acting elements, specifically a single-stranded origin and a double-stranded origin, which are characteristic of rolling-circle-replication plasmids. Experiments with pRE8424 derivatives carrying a mutated single-stranded origin sequence showed that single-stranded DNA intermediates accumulated in the cells because of inefficient conversion from single-stranded DNA to double

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Hingston, Patricia, Thomas Brenner, Lisbeth Truelstrup Hansen, and Siyun Wang. "Comparative Analysis of Listeria monocytogenes Plasmids and Expression Levels of Plasmid-Encoded Genes during Growth under Salt and Acid Stress Conditions." Toxins 11, no. 7 (2019): 426. http://dx.doi.org/10.3390/toxins11070426.

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Listeria monocytogenes strains are known to harbour plasmids that confer resistance to sanitizers, heavy metals, and antibiotics; however, very little research has been conducted into how plasmids may influence L. monocytogenes’ ability to tolerate food-related stresses. To investigate this, a library (n = 93) of L. monocytogenes plasmid sequences were compared. Plasmid sequences were divided into two groups (G1 and G2) based on a repA phylogeny. Twenty-six unique plasmid types were observed, with 13 belonging to each of the two repA-based groups. G1 plasmids were significantly (p < 0.05) s

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Cook, L. C., and G. M. Dunny. "Effects of Biofilm Growth on Plasmid Copy Number and Expression of Antibiotic Resistance Genes in Enterococcus faecalis." Antimicrobial Agents and Chemotherapy 57, no. 4 (2013): 1850–56. http://dx.doi.org/10.1128/aac.02010-12.

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ABSTRACTBiofilm growth causes increased average plasmid copy number as well as increased copy number heterogeneity inEnterococcus faecaliscells carrying plasmid pCF10. In this study, we examined whether biofilm growth affected the copy number and expression of antibiotic resistance determinants for several plasmids with diverse replication systems. Four differentE. faecalisplasmids, unrelated to pCF10, demonstrated increased copy number in biofilm cells. In biofilm cells, we also observed increased transcription of antibiotic resistance genes present on these plasmids. The increase in plasmid

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8

O'Flaherty, Sarah, and Todd R. Klaenhammer. "Multivalent Chromosomal Expression of the Clostridium botulinum Serotype A Neurotoxin Heavy-Chain Antigen and the Bacillus anthracis Protective Antigen in Lactobacillus acidophilus." Applied and Environmental Microbiology 82, no. 20 (2016): 6091–101. http://dx.doi.org/10.1128/aem.01533-16.

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ABSTRACTClostridium botulinumandBacillus anthracisproduce potent toxins that cause severe disease in humans. New and improved vaccines are needed for both of these pathogens. For mucosal vaccine delivery using lactic acid bacteria, chromosomal expression of antigens is preferred over plasmid-based expression systems, as chromosomal expression circumvents plasmid instability and the need for antibiotic pressure. In this study, we constructed three strains ofLactobacillus acidophilusNCFM expressing from the chromosome (i) the nontoxic host receptor-binding domain of the heavy chain ofClostridium

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9

Kitten, T., and A. G. Barbour. "The relapsing fever agent Borrelia hermsii has multiple copies of its chromosome and linear plasmids." Genetics 132, no. 2 (1992): 311–24. http://dx.doi.org/10.1093/genetics/132.2.311.

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Abstract Borrelia hermsii, a spirochete which causes relapsing fever in humans and other mammals, eludes the immune response by antigenic variation of the "Vmp" proteins. This occurs by replacement of an expressed vmp gene with a copy of a silent vmp gene. Silent and expressed vmp genes are located on separate linear plasmids. To further characterize vmp recombination, copy numbers were determined for two linear plasmids and for the 1-megabase chromosome by comparing hybridization of probes to native DNA with hybridization to recombinant plasmids containing borrelial DNA. Plasmid copy numbers

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10

Madriz, Xochil, Máximo B. Martínez, Mario A. Rodríguez, et al. "Expression in fibroblasts and in live animals of Entamoeba histolytica polypeptides EhCP112 and EhADH112." Microbiology 150, no. 5 (2004): 1251–60. http://dx.doi.org/10.1099/mic.0.26938-0.

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EhCPADH is an immunogenic, heterodimeric protein that is formed by EhCP112 (cysteine protease) and EhADH112 (adhesin), polypeptides involved in Entamoeba histolytica's cytopathic effect, target-cell adherence and phagocytosis. The EhCPADH complex is located in the plasma membrane and cytoplasmic vacuoles. Here, the independent expression of EhCP112 and EhADH112 in fibroblasts and hamsters was analysed. Also investigated was the immunological response in animals independently inoculated with plasmid pcDNA-Ehcp112, which carries the complete cysteine protease-encoding gene, or with plasmid pcDNA

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11

Philip, R., E. Brunette, L. Kilinski, et al. "Efficient and sustained gene expression in primary T lymphocytes and primary and cultured tumor cells mediated by adeno-associated virus plasmid DNA complexed to cationic liposomes." Molecular and Cellular Biology 14, no. 4 (1994): 2411–18. http://dx.doi.org/10.1128/mcb.14.4.2411.

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We have used cationic liposomes to facilitate adeno-associated virus (AAV) plasmid transfections of primary and cultured cell types. AAV plasmid DNA complexed with liposomes showed levels of expression several fold higher than those of complexes with standard plasmids. In addition, long-term expression (> 30 days) of the gene, unlike the transient expression demonstrated by typical liposome-mediated transfection with standard plasmids, was observed. Southern analysis of chromosomal DNA further substantiated the hypothesis that the long-term expression was due to the presence of the transgen

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Philip, R., E. Brunette, L. Kilinski, et al. "Efficient and sustained gene expression in primary T lymphocytes and primary and cultured tumor cells mediated by adeno-associated virus plasmid DNA complexed to cationic liposomes." Molecular and Cellular Biology 14, no. 4 (1994): 2411–18. http://dx.doi.org/10.1128/mcb.14.4.2411-2418.1994.

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We have used cationic liposomes to facilitate adeno-associated virus (AAV) plasmid transfections of primary and cultured cell types. AAV plasmid DNA complexed with liposomes showed levels of expression several fold higher than those of complexes with standard plasmids. In addition, long-term expression (> 30 days) of the gene, unlike the transient expression demonstrated by typical liposome-mediated transfection with standard plasmids, was observed. Southern analysis of chromosomal DNA further substantiated the hypothesis that the long-term expression was due to the presence of the transgen

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13

Fonzi, W. A., and P. S. Sypherd. "Expression of the gene for ornithine decarboxylase of Saccharomyces cerevisiae in Escherichia coli." Molecular and Cellular Biology 5, no. 1 (1985): 161–66. http://dx.doi.org/10.1128/mcb.5.1.161.

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Diploid cells of Saccharomyces cerevisiae homozygous for the spe1A mutation, which eliminates ornithine decarboxylase activity, were found to sporulate at a greatly reduced frequency in the absence of polyamines. Plasmids which complement the spe1A mutation were isolated by their ability to restore sporulation competence to these cells. Three distinct plasmids were isolated. Each plasmid insert overlapped the same 8.0-kilobase region, and each plasmid restored ornithine decarboxylase activity to spe1A mutants. These plasmids also conferred ornithine decarboxylase activity to Escherichia coli E

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Fonzi, W. A., and P. S. Sypherd. "Expression of the gene for ornithine decarboxylase of Saccharomyces cerevisiae in Escherichia coli." Molecular and Cellular Biology 5, no. 1 (1985): 161–66. http://dx.doi.org/10.1128/mcb.5.1.161-166.1985.

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Diploid cells of Saccharomyces cerevisiae homozygous for the spe1A mutation, which eliminates ornithine decarboxylase activity, were found to sporulate at a greatly reduced frequency in the absence of polyamines. Plasmids which complement the spe1A mutation were isolated by their ability to restore sporulation competence to these cells. Three distinct plasmids were isolated. Each plasmid insert overlapped the same 8.0-kilobase region, and each plasmid restored ornithine decarboxylase activity to spe1A mutants. These plasmids also conferred ornithine decarboxylase activity to Escherichia coli E

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15

Sayeed, Sameera, Lucretia Reaves, Lyndsay Radnedge, and Stuart Austin. "The Stability Region of the Large Virulence Plasmid ofShigella flexneri Encodes an Efficient Postsegregational Killing System." Journal of Bacteriology 182, no. 9 (2000): 2416–21. http://dx.doi.org/10.1128/jb.182.9.2416-2421.2000.

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ABSTRACT The large virulence plasmid pMYSH6000 of Shigella flexneri contains a determinant that is highly effective in stabilizing otherwise unstable plasmids in Escherichia coli. Expression of two small contiguous genes, mvpAand mvpT (formerly termed STBORF1 and STBORF2), was shown to be sufficient for stability. Mutations in mvpT abolished plasmid stability, and plasmids expressing only mvpT killed the cells unless mvpA was supplied from a separate plasmid or from the host chromosome. When replication of a plasmid carrying the minimal mvp region was blocked, growth of the culture stopped aft

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Yang, Jun, Hai-Hong Wang, Yaoyao Lu, et al. "A ProQ/FinO family protein involved in plasmid copy number control favours fitness of bacteria carrying mcr-1-bearing IncI2 plasmids." Nucleic Acids Research 49, no. 7 (2021): 3981–96. http://dx.doi.org/10.1093/nar/gkab149.

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Abstract The plasmid-encoded colistin resistance gene mcr-1 challenges the use of polymyxins and poses a threat to public health. Although IncI2-type plasmids are the most common vector for spreading the mcr-1 gene, the mechanisms by which these plasmids adapt to host bacteria and maintain resistance genes remain unclear. Herein, we investigated the regulatory mechanism for controlling the fitness cost of an IncI2 plasmid carrying mcr-1. A putative ProQ/FinO family protein encoded by the IncI2 plasmid, designated as PcnR (plasmid copy number repressor), balances the mcr-1 expression and bacter

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17

Zhang, Hongyang, Mingding Chang, Xiaochen Zhang, et al. "Functional Identification and Evolutionary Analysis of Two Novel Plasmids Mediating Quinolone Resistance in Proteus vulgaris." Microorganisms 8, no. 7 (2020): 1074. http://dx.doi.org/10.3390/microorganisms8071074.

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Plasmid-mediated quinolone resistance (PMQR) remains one of the main mechanisms of bacterial quinolone resistance and plays an important role in the transmission of antibiotic resistance genes (ARGs). In this study, two novel plasmids, p3M-2A and p3M-2B, which mediate quinolone resistance in Proteus vulgaris strain 3M (P3M) were identified. Of these, only p3M-2B appeared to be a qnrD-carrying plasmid. Both p3M-2A and p3M-2B could be transferred into Escherichia coli, and the latter caused a twofold change in ciprofloxacin resistance, according to the measured minimum inhibitory concentration (

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Haryanto, Aris, Hevi Wihadmadyatami, and Nastiti Wijayanti. "In vitro expression of the recombinant fusion protein of Newcastle disease virus from local Indonesian isolates by using a cell-free protein expression system." Indonesian Journal of Biotechnology 25, no. 2 (2020): 69. http://dx.doi.org/10.22146/ijbiotech.54703.

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The aim of this work was the in vitro expression of the recombinant fusion (F) protein of Newcastle disease virus (NDV). The pBT7-N-His-Fusion-NDV expression plasmid which carries the recombinant F protein encoding gene from local Indonesian isolates, was prepared and transformed into E. coli BL21 (DE3). To detect bacterial colonies carrying the recombinant plasmid, a restriction endonuclease analysis was performed using the EcoRI restriction endonuclease. These results showed that the pBT-N-His-Fusion-NDV plasmid was successfully isolated with a size of 4.601 bp, and three recombinant plasmid

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Hausjell, Johanna, Regina Kutscha, Jeannine D. Gesson, Daniela Reinisch, and Oliver Spadiut. "The Effects of Lactose Induction on a Plasmid-Free E. coli T7 Expression System." Bioengineering 7, no. 1 (2020): 8. http://dx.doi.org/10.3390/bioengineering7010008.

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Recombinant production of pharmaceutical proteins like antigen binding fragments (Fabs) in the commonly-used production host Escherichia coli presents several challenges. The predominantly-used plasmid-based expression systems exhibit the drawback of either excessive plasmid amplification or plasmid loss over prolonged cultivations. To improve production, efforts are made to establish plasmid-free expression, ensuring more stable process conditions. Another strategy to stabilize production processes is lactose induction, leading to increased soluble product formation and cell fitness, as shown

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Quinlan, M. P., and D. M. Knipe. "Stimulation of expression of a herpes simplex virus DNA-binding protein by two viral functions." Molecular and Cellular Biology 5, no. 5 (1985): 957–63. http://dx.doi.org/10.1128/mcb.5.5.957.

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We examined the expression and localization of herpesvirus proteins in monkey cells transfected with recombinant plasmids containing herpes simplex virus (HSV) DNA sequences. Low levels of expression of the major HSV DNA-binding protein ICP8 were observed when ICP8-encoding plasmids were introduced into cells alone. ICP8 expression was greatly increased when a recombinant plasmid encoding the HSV alpha (immediate-early) ICP4 and ICP0 genes was transfected with the ICP8 gene. Deletion and subcloning analysis indicated that two separate functions capable of stimulating ICP8 expression were encod

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Quinlan, M. P., and D. M. Knipe. "Stimulation of expression of a herpes simplex virus DNA-binding protein by two viral functions." Molecular and Cellular Biology 5, no. 5 (1985): 957–63. http://dx.doi.org/10.1128/mcb.5.5.957-963.1985.

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We examined the expression and localization of herpesvirus proteins in monkey cells transfected with recombinant plasmids containing herpes simplex virus (HSV) DNA sequences. Low levels of expression of the major HSV DNA-binding protein ICP8 were observed when ICP8-encoding plasmids were introduced into cells alone. ICP8 expression was greatly increased when a recombinant plasmid encoding the HSV alpha (immediate-early) ICP4 and ICP0 genes was transfected with the ICP8 gene. Deletion and subcloning analysis indicated that two separate functions capable of stimulating ICP8 expression were encod

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Khanna-Gupta, Arati, Theresa Zibello, Sarah Kolla, Ellis J. Neufeld, and Nancy Berliner. "CCAAT Displacement Protein (CDP/cut) Recognizes a Silencer Element Within the Lactoferrin Gene Promoter." Blood 90, no. 7 (1997): 2784–95. http://dx.doi.org/10.1182/blood.v90.7.2784.

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Abstract Expression of neutrophil secondary granule protein (SGP) genes is coordinately regulated at the transcriptional level, and is disrupted in specific granule deficiency and leukemia. We analyzed the regulation of SGP gene expression by luciferase reporter gene assays using the lactoferrin (LF) promoter. Reporter plasmids were transiently transfected into non–LF-expressing hematopoietic cell lines. Luciferase activity was detected from reporter plasmids containing basepair (bp) −387 to bp −726 of the LF promoter, but not in a −916-bp plasmid. Transfection of a −916-bp plasmid into a LF-e

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Khanna-Gupta, Arati, Theresa Zibello, Sarah Kolla, Ellis J. Neufeld, and Nancy Berliner. "CCAAT Displacement Protein (CDP/cut) Recognizes a Silencer Element Within the Lactoferrin Gene Promoter." Blood 90, no. 7 (1997): 2784–95. http://dx.doi.org/10.1182/blood.v90.7.2784.2784_2784_2795.

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Expression of neutrophil secondary granule protein (SGP) genes is coordinately regulated at the transcriptional level, and is disrupted in specific granule deficiency and leukemia. We analyzed the regulation of SGP gene expression by luciferase reporter gene assays using the lactoferrin (LF) promoter. Reporter plasmids were transiently transfected into non–LF-expressing hematopoietic cell lines. Luciferase activity was detected from reporter plasmids containing basepair (bp) −387 to bp −726 of the LF promoter, but not in a −916-bp plasmid. Transfection of a −916-bp plasmid into a LF-expressing

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Loftie-Eaton, Wesley, and Douglas E. Rawlings. "Comparative Biology of Two Natural Variants of the IncQ-2 Family Plasmids, pRAS3.1 and pRAS3.2." Journal of Bacteriology 191, no. 20 (2009): 6436–46. http://dx.doi.org/10.1128/jb.00864-09.

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ABSTRACT Plasmids pRAS3.1 and pRAS3.2 are two closely related, natural variants of the IncQ-2 plasmid family that have identical plasmid backbones except for two differences. Plasmid pRAS3.1 has five 6-bp repeat sequences in the promoter region of the mobB gene and four 22-bp iterons in its oriV region, whereas pRAS3.2 has only four 6-bp repeats and three 22-bp iterons. Plasmid pRAS3.1 was found to have a higher copy number than pRAS3.2, and we show that the extra 6-bp repeat results in an increase in mobB and downstream mobA/repB expression. Placement of repB (primase) behind an arabinose-ind

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Zimmermann, Alice, David Hercher, Benedikt Regner, et al. "Evaluation of BMP-2 Minicircle DNA for Enhanced Bone Engineering and Regeneration." Current Gene Therapy 20, no. 1 (2020): 55–63. http://dx.doi.org/10.2174/1566523220666200427121350.

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Background: To date, the significant osteoinductive potential of bone morphogenetic protein 2 (BMP-2) non-viral gene therapy cannot be fully exploited therapeutically. This is mainly due to weak gene delivery and brief expression peaks restricting the therapeutic effect. Objective: Our objective was to test the application of minicircle DNA, allowing prolonged expression potential. It offers notable advantages over conventional plasmid DNA. The lack of bacterial sequences and the resulting reduction in size, enables safe usage and improved performance for tissue regeneration. Methods: We inser

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Soares, Alexandra, Luciana C. Gomes, Gabriel A. Monteiro, and Filipe J. Mergulhão. "The Influence of Nutrient Medium Composition on Escherichia coli Biofilm Development and Heterologous Protein Expression." Applied Sciences 11, no. 18 (2021): 8667. http://dx.doi.org/10.3390/app11188667.

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In the present study, the effects of different nutrient media on the development of Escherichia coli biofilms and the production of a heterologous protein were examined. E. coli JM109(DE3) cells transformed with pFM23 plasmid carrying the gene for enhanced green fluorescent protein (eGFP) expression were used. Cells were grown in two different culture media, Lysogenic Broth (LB) and M9ZB, in a flow cell system for 10 days. Epifluorescence microscopy, fluorimetry, and a high-performance liquid chromatography (HPLC) method based on hydrophobic interaction chromatography (HIC) were used to assess

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Rajanna, Chythanya, Tamara Revazishvili, Mohammed H. Rashid, et al. "Characterization of pPCP1 Plasmids inYersinia pestisStrains Isolated from the Former Soviet Union." International Journal of Microbiology 2010 (2010): 1–9. http://dx.doi.org/10.1155/2010/760819.

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Complete sequences of 9.5-kb pPCP1 plasmids in threeYersinia pestisstrains from the former Soviet Union (FSU) were determined and compared with those of pPCP1 plasmids in three well-characterized, non-FSUY. pestisstrains (KIM, CO92, and 91001). Two of the FSU plasmids were from strains C2614 and C2944, isolated from plague foci in Russia, and one plasmid was from strain C790 from Kyrgyzstan. Sequence analyses identified four sequence types among the six plasmids. The pPCP1 plasmids in the FSU strains were most genetically related to the pPCP1 plasmid in the KIM strain and least related to the

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Adamczyk, Małgorzata, and Grazyna Jagura-Burdzy. "Spread and survival of promiscuous IncP-1 plasmids." Acta Biochimica Polonica 50, no. 2 (2003): 425–53. http://dx.doi.org/10.18388/abp.2003_3696.

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Plasmids classified to the IncP-1 incompatibility group belong to the most stably maintained mobile elements among low copy number plasmids known to date. The remarkable persistence is achieved by various tightly controlled stability mechanisms like active partitioning, efficient conjugative transfer system, killing of plasmid-free segregants and multimer resolution. The unique feature of IncP-1 plasmids is the central control operon coding for global regulators which control the expression of genes involved in vegetative replication, stable maintenance and conjugative transfer. The multivalen

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Khanna-Gupta, Arati, Theresa Zibello, Carl Simkevich, Alan G. Rosmarin, and Nancy Berliner. "Sp1 and C/EBP are necessary to activate the lactoferrin gene promoter during myeloid differentiation." Blood 95, no. 12 (2000): 3734–41. http://dx.doi.org/10.1182/blood.v95.12.3734.

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Abstract In this study, we sought to identify factors responsible for the positive modulation of lactoferrin (LF), a neutrophil-specific, secondary-granule protein gene. Initial reporter gene transfection assays indicated that the first 89 base pairs of the LF promoter are capable of directing myeloid-specific LF gene expression. The presence of a C/EBP site flanked by 2 Sp1 sites within this segment of the LF promoter prompted us to investigate the possible role of these sites in LF expression. Cotransfection studies of LF-89luc plasmid with increasing concentrations of a C/EBP expression ve

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Khanna-Gupta, Arati, Theresa Zibello, Carl Simkevich, Alan G. Rosmarin, and Nancy Berliner. "Sp1 and C/EBP are necessary to activate the lactoferrin gene promoter during myeloid differentiation." Blood 95, no. 12 (2000): 3734–41. http://dx.doi.org/10.1182/blood.v95.12.3734.012k27_3734_3741.

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In this study, we sought to identify factors responsible for the positive modulation of lactoferrin (LF), a neutrophil-specific, secondary-granule protein gene. Initial reporter gene transfection assays indicated that the first 89 base pairs of the LF promoter are capable of directing myeloid-specific LF gene expression. The presence of a C/EBP site flanked by 2 Sp1 sites within this segment of the LF promoter prompted us to investigate the possible role of these sites in LF expression. Cotransfection studies of LF-89luc plasmid with increasing concentrations of a C/EBP expression vector in m

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McGuffie, Matthew J., and Jeffrey E. Barrick. "pLannotate: engineered plasmid annotation." Nucleic Acids Research 49, W1 (2021): W516—W522. http://dx.doi.org/10.1093/nar/gkab374.

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Abstract Engineered plasmids are widely used in the biological sciences. Since many plasmids contain DNA sequences that have been reused and remixed by researchers for decades, annotation of their functional elements is often incomplete. Missing information about the presence, location, or precise identity of a plasmid feature can lead to unintended consequences or failed experiments. Many engineered plasmids contain sequences—such as recombinant DNA from all domains of life, wholly synthetic DNA sequences, and engineered gene expression elements—that are not predicted by microbial genome anno

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Liu, Baomo, Lili Shui, Kai Zhou, et al. "Impact of Plasmid-Encoded H-NS–like Protein on blaNDM-1-Bearing IncX3 Plasmid in Escherichia coli." Journal of Infectious Diseases 221, Supplement_2 (2020): S229—S236. http://dx.doi.org/10.1093/infdis/jiz567.

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Abstract Background This study was performed to assess the role of the histone-like nucleoid-structuring (H-NS)–like protein, carried by blaNDM-1-encoding IncX3-type plasmids, in the dissemination of IncX3 plasmids. Methods The blaNDM-1-encoding IncX3 plasmids were analyzed using southern blot, conjugation, and competition assays. Virulence was evaluated with a Galleria mellonella infection model. An hns-knockout IncX3 plasmid was also constructed to identify the functions of plasmid-borne H-NS–like protein in Escherichia coli. Results The assasys detected blaNDM-1-encoding IncX3-type plasmids

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Wu, Yuntao, Ge Liu, and Eric B. Carstens. "Replication, Integration, and Packaging of Plasmid DNA following Cotransfection with Baculovirus Viral DNA." Journal of Virology 73, no. 7 (1999): 5473–80. http://dx.doi.org/10.1128/jvi.73.7.5473-5480.1999.

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ABSTRACT Infection-dependent replication assays have been used to identify numerous putative origins of baculovirus replication. However, plasmid DNA, when cotransfected into insect cells with Autographa californica multinucleocapsid nucleopolyhedrovirus (AcMNPV) DNA, replicates independently of any viral sequence in cis (11). Cotransfection of transfer plasmids and baculovirus DNA is a common procedure used in generating recombinant viruses and in measuring the level of gene expression in transient-expression assays. We have examined the fate of a series of vector plasmids in cotransfection e

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Grimm, Dorothee, Abdallah F. Elias, Kit Tilly, and Patricia A. Rosa. "Plasmid Stability during In Vitro Propagation of Borrelia burgdorferi Assessed at a Clonal Level." Infection and Immunity 71, no. 6 (2003): 3138–45. http://dx.doi.org/10.1128/iai.71.6.3138-3145.2003.

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ABSTRACT Borrelia burgdorferi causes Lyme disease in humans. The genome of the sequenced type strain B31 MI consists of a linear chromosome, 12 linear plasmids, and 9 circular plasmids. Previous studies by other investigators indicated that some of these plasmids are essential for the survival of the spirochetes in vivo but not in vitro. We have studied plasmid stability during in vitro growth at 23 and 35°C, conditions that approximate the temperatures of the tick vector and the mammalian host, respectively. Starting with two clones that have all 21 plasmids, we investigated plasmid maintenan

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Saksena, Nitin K., and Nicole Truffaut. "Cloning of tetracycline-resistance genes from various strains of Clostridium perfringens and expression in Escherichia coli." Canadian Journal of Microbiology 38, no. 3 (1992): 215–21. http://dx.doi.org/10.1139/m92-036.

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One hundred strains of Clostridium perfringens and 52 strains of other Clostridia of human and animal origins were screened for tetracycline resistance. Fifty-six strains were resistant to tetracycline in the C. perfringens group. Ten strains were selected for their high level of resistance. In all of them, the tetracycline-resistance genes were found to be residing in large plasmids of about 50 kb, all showing homologies. Several tetracycline-resistance genes from plasmids of various strains of C. perfringens were cloned in plasmid pUC19 and the resistance was expressed in Escherichia coli. H

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Alwine, J. C. "Transient gene expression control: effects of transfected DNA stability and trans-activation by viral early proteins." Molecular and Cellular Biology 5, no. 5 (1985): 1034–42. http://dx.doi.org/10.1128/mcb.5.5.1034.

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The effects of trans-acting factors and transfected DNA stability on promoter activity were examined with chloramphenicol acetyl transferase (CAT) transient expression analysis. With cotransfection into CV-1P and HeLa cells, simian virus 40 T antigen, adenovirus E1a, and herpes-virus IE proteins were compared for their ability to trans-activate a variety of eucaryotic promoters constructed into CAT plasmids. T antigen and the IE protein were promiscuous activators of all the promoters tested [the simian virus 40 late promoter, the adenovirus E3 promoter, the alpha 2(I) collagen promoter, and t

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37

Alwine, J. C. "Transient gene expression control: effects of transfected DNA stability and trans-activation by viral early proteins." Molecular and Cellular Biology 5, no. 5 (1985): 1034–42. http://dx.doi.org/10.1128/mcb.5.5.1034-1042.1985.

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The effects of trans-acting factors and transfected DNA stability on promoter activity were examined with chloramphenicol acetyl transferase (CAT) transient expression analysis. With cotransfection into CV-1P and HeLa cells, simian virus 40 T antigen, adenovirus E1a, and herpes-virus IE proteins were compared for their ability to trans-activate a variety of eucaryotic promoters constructed into CAT plasmids. T antigen and the IE protein were promiscuous activators of all the promoters tested [the simian virus 40 late promoter, the adenovirus E3 promoter, the alpha 2(I) collagen promoter, and t

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Zielenkiewicz, Urszula, and Piotr Cegłowski. "The Toxin-Antitoxin System of the Streptococcal Plasmid pSM19035." Journal of Bacteriology 187, no. 17 (2005): 6094–105. http://dx.doi.org/10.1128/jb.187.17.6094-6105.2005.

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ABSTRACT pSM19035 of the pathogenic bacterium Streptococcus pyogenes is a low-copy-number plasmid carrying erythromycin resistance, stably maintained in a broad range of gram-positive bacteria. We show here that the ω-ε-ζ operon of this plasmid constitutes a novel proteic plasmid addiction system in which the ε and ζ genes encode an antitoxin and toxin, respectively, while ω plays an autoregulatory function. Expression of toxin Zeta is bactericidal for the gram-positive Bacillus subtilis and bacteriostatic for the gram-negative Escherichia coli. The toxic effects of ζ gene expression in both b

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McNeill, Hesta V., Katharine A. Sinha, Carlos E. Hormaeche, Jeong Jin Lee, and C. M. Anjam Khan. "Development of a Nonantibiotic Dominant Marker for Positively Selecting Expression Plasmids in MultivalentSalmonella Vaccines." Applied and Environmental Microbiology 66, no. 3 (2000): 1216–19. http://dx.doi.org/10.1128/aem.66.3.1216-1219.2000.

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ABSTRACT We report the novel application of a herbicide-resistance-based dominant marker for the positive selection of expression plasmids inSalmonella serovar vaccines. The β-lactamase gene of the plasmid pTETnir15, which expresses fragment C of tetanus toxin (TetC), has been replaced with the bar gene marker. The new plasmid pBAT1 can be positively selected in vitro within Salmonellaserovars in the presence of the herbicidedl-phosphinothricin. The expression of TetC remains unaltered, and the Salmonella enterica serovar Typhimurium vaccine strain is stable and immunogenic in vivo.

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Bergsveinson, Jordyn, Nina Baecker, Vanessa Pittet, and Barry Ziola. "Role of Plasmids in Lactobacillus brevis BSO 464 Hop Tolerance and Beer Spoilage." Applied and Environmental Microbiology 81, no. 4 (2014): 1234–41. http://dx.doi.org/10.1128/aem.02870-14.

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ABSTRACTSpecific isolates of lactic acid bacteria (LAB) can grow in the harsh beer environment, thus posing a threat to brew quality and the economic success of breweries worldwide. Plasmid-localized genes, such ashorA,horC, andhitA, have been suggested to confer hop tolerance, a trait required for LAB survival in beer. The presence and expression of these genes among LAB, however, do not universally correlate with the ability to grow in beer. Genome sequencing of the virulent beer spoilage organismLactobacillus brevisBSO 464 revealed the presence of eight plasmids, with plasmids 1, 2, and 3 c

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Hubberstey, Andrew V., and Alan G. Wildeman. "Use of interplasmid recombination to generate stable selectable markers for yeast transformation: application to studies of actin gene control." Genome 33, no. 5 (1990): 696–706. http://dx.doi.org/10.1139/g90-105.

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A plasmid recombination system has been developed that relies upon interplasmid exchanges for yeast cell viability. Two types of plasmids, one carrying the LEU2 allele inserted within yeast actin gene sequences and the other carrying 2-μm plasmid DNA and an intact actin gene, were constructed. Neither plasmid alone yielded transformants in the haploid Leu− strain AH22, but when cotransformed, a number of colonies were obtained. Southern blot analysis revealed that transformants arose because of recombination events within the homologous actin sequences that transferred the LEU2 gene to the act

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Qin, Zhongjun, Meijuan Shen, and Stanley N. Cohen. "Identification and Characterization of a pSLA2 Plasmid Locus Required for Linear DNA Replication and Circular Plasmid Stable Inheritance in Streptomyces lividans." Journal of Bacteriology 185, no. 22 (2003): 6575–82. http://dx.doi.org/10.1128/jb.185.22.6575-6582.2003.

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ABSTRACT Streptomyces linear plasmids and linear chromosomes can replicate also in a circular form when their telomeres are deleted. The 17-kb linear plasmid pSLA2 has been a useful model in studies of such replicons. Here we report that the minimal origin initiating replication of pSLA2-derived plasmids as circular molecules cannot propagate these plasmids in a linear mode unless they also contain a novel plasmid-encoded locus, here named rlrA (required for linear replication). In contrast with the need for rlrA to accomplish replication of telomere-containing linear plasmids, expression of r

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Giguère, Steeve, Mary K. Hondalus, Julie A. Yager, Patricia Darrah, David M. Mosser, and John F. Prescott. "Role of the 85-Kilobase Plasmid and Plasmid-Encoded Virulence-Associated Protein A in Intracellular Survival and Virulence of Rhodococcus equi." Infection and Immunity 67, no. 7 (1999): 3548–57. http://dx.doi.org/10.1128/iai.67.7.3548-3557.1999.

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ABSTRACT Rhodococcus equi is a facultative intracellular pathogen of macrophages and a cause of pneumonia in young horses (foals) and immunocompromised people. Isolates of R. equi from pneumonic foals typically contain large, 85- or 90-kb plasmids encoding a highly immunogenic virulence-associated protein (VapA). The objective of this study was to determine the role of the 85-kb plasmid and VapA in the intracellular survival and virulence of R. equi. Clinical isolates containing the plasmid and expressing VapA efficiently replicated within mouse macrophages in vitro, while plasmid-cured deriva

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Durand, Romain, Kévin T. Huguet, Nicolas Rivard, Nicolas Carraro, Sébastien Rodrigue, and Vincent Burrus. "Crucial role of Salmonella genomic island 1 master activator in the parasitism of IncC plasmids." Nucleic Acids Research 49, no. 14 (2021): 7807–24. http://dx.doi.org/10.1093/nar/gkab204.

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Abstract IncC conjugative plasmids and the multiple variants of Salmonella Genomic Island 1 (SGI1) are two functionally interacting families of mobile genetic elements commonly associated with multidrug resistance in the Gammaproteobacteria. SGI1 and its siblings are specifically mobilised in trans by IncC conjugative plasmids. Conjugative transfer of IncC plasmids is activated by the plasmid-encoded master activator AcaCD. SGI1 carries five AcaCD-responsive promoters that drive the expression of genes involved in its excision, replication, and mobilisation. SGI1 encodes an AcaCD homologue, th

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Ginns, C. A., M. L. Benham, L. M. Adams, et al. "Colonization of the Respiratory Tract by a Virulent Strain of Avian Escherichia coli Requires Carriage of a Conjugative Plasmid." Infection and Immunity 68, no. 3 (2000): 1535–41. http://dx.doi.org/10.1128/iai.68.3.1535-1541.2000.

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ABSTRACT The E3 strain of E. coli was isolated in an outbreak of respiratory disease in broiler chickens, and experimental aerosol exposure of chickens to this strain induced disease similar to that seen in the field. In order to establish whether the virulent phenotype of this strain was associated with carriage of particular plasmids, four plasmid-cured derivatives, each lacking two or more of the plasmids carried by the wild-type strain, were assessed for virulence. Virulence was found to be associated with one large plasmid, pVM01. Plasmid pVM01 was marked by introduction of the transposon

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Kues, W. A., K. Iqbal, B. Barg-Kues, and H. Niemann. "305 REPROGRAMMING EVENTS IN EARLY BOVINE AND MURINE EMBRYOS ARE MIRRORED BY PLASMID-ENCODED MARKER CONSTRUCTS." Reproduction, Fertility and Development 21, no. 1 (2009): 249. http://dx.doi.org/10.1071/rdv21n1ab305.

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Episomal plasmids have emerged as useful tools to achieve stable transgenesis in mammalian cell cultures. Here, the suitability of scaffold/matrix attachment region (S/MAR) carrying episomal plasmids and conventional plasmids for the generation of transgenic murine and bovine embryos was assessed. Bovine zygotes were produced from slaughterhouse ovaries, and murine zygotes were isolated from superovulated and mated NMRI females. Zygote stages were microinjected with approximately 10 pl of plasmid solution. The S/MAR encoding plasmids pEPI or minicircle preparations (gift of J. Bode, Braunschwe

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47

Wonohadi, Elisawati, and Mariana Wahyudi. "KLONING GEN carB Salmonella typhi MENGGUNAKAN VEKTOR EKSPRESI pET-16b dan SEL INANG Escherichia coli JM109." Sains & Teknologi 3, no. 3 (2019): 57. http://dx.doi.org/10.24123/jst.v3i3.2289.

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The effort of cloning the Lister strain of Salmonella typhi (NCTC 786, BCC 712) carB gene using pET-16b expression vector and E. coli JM109 as host cell has been done. The carB gene and the pET-16b expression vector were both prepared from their recombinant plasmid digested using BamHI and NdeI as restriction enzymes. The pG-carB-11-ST recombinant plasmid was isolated from Escherichia coli XL10(pG-carB-11-ST) and the pET-carA-ST recombinant plasmid was from Escherichia coli DH5α(pET-carA-ST). After being ligated (in a ratio of vector:gene of 1:3, 1:5 and 1:6) in the presence of T4 Bacteriofage

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Schuster, Layla A., and Christopher R. Reisch. "A plasmid toolbox for controlled gene expression across the Proteobacteria." Nucleic Acids Research 49, no. 12 (2021): 7189–202. http://dx.doi.org/10.1093/nar/gkab496.

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Abstract Controlled gene expression is fundamental for the study of gene function and our ability to engineer bacteria. However, there is currently no easy-to-use genetics toolbox that enables controlled gene expression in a wide range of diverse species. To facilitate the development of genetics systems in a fast, easy, and standardized manner, we constructed and tested a plasmid assembly toolbox that will enable the identification of well-regulated promoters in many Proteobacteria and potentially beyond. Each plasmid is composed of four categories of genetic parts (i) the origin of replicati

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Kloos, Julia, João A. Gama, Joachim Hegstad, Ørjan Samuelsen, and Pål J. Johnsen. "Piggybacking on Niche Adaptation Improves the Maintenance of Multidrug-Resistance Plasmids." Molecular Biology and Evolution 38, no. 8 (2021): 3188–201. http://dx.doi.org/10.1093/molbev/msab091.

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Abstract The persistence of plasmids in bacterial populations represents a puzzling evolutionary problem with serious clinical implications due to their role in the ongoing antibiotic resistance crisis. Recently, major advancements have been made toward resolving this “plasmid paradox” but mainly in a nonclinical context. Here, we propose an additional explanation for the maintenance of multidrug-resistance plasmids in clinical Escherichia coli strains. After coevolving two multidrug-resistance plasmids encoding resistance to last resort carbapenems with an extraintestinal pathogenic E. coli s

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Bao, G. Y., K. Y. Lu, S. F. Cui, and L. Xu. "DKK1 eukaryotic expression plasmid and expression product identification." Genetics and Molecular Research 14, no. 2 (2015): 6312–18. http://dx.doi.org/10.4238/2015.june.11.5.

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