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Journal articles on the topic 'Full-length clone'

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Published: 7 June 2025

Last updated: 2 August 2025

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1

Wolf, D., N. Harris, N. Goldfinger, and V. Rotter. "Isolation of a full-length mouse cDNA clone coding for an immunologically distinct p53 molecule." Molecular and Cellular Biology 5, no. 1 (1985): 127–32. http://dx.doi.org/10.1128/mcb.5.1.127-132.1985.

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Transfection of a cloned p53 gene into a p53 nonproducer Abelson murine leukemia virus-transformed cell line, L12, reconstituted p53 expression. The protein expressed in these cells was indistinguishable from that naturally expressed in p53 producer tumor cells. Conversely, p53 protein expressed in L12-derived clones that were established by transfection with a full-length p53 cDNA clone (pM8) exhibited a discrete immunological form. Immunoprecipitation of p53 with a panel of monoclonal anti-p53 antibodies showed that L12-derived clones that were transfected with the genomic p53 clone containe

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2

Wolf, D., N. Harris, N. Goldfinger, and V. Rotter. "Isolation of a full-length mouse cDNA clone coding for an immunologically distinct p53 molecule." Molecular and Cellular Biology 5, no. 1 (1985): 127–32. http://dx.doi.org/10.1128/mcb.5.1.127.

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Transfection of a cloned p53 gene into a p53 nonproducer Abelson murine leukemia virus-transformed cell line, L12, reconstituted p53 expression. The protein expressed in these cells was indistinguishable from that naturally expressed in p53 producer tumor cells. Conversely, p53 protein expressed in L12-derived clones that were established by transfection with a full-length p53 cDNA clone (pM8) exhibited a discrete immunological form. Immunoprecipitation of p53 with a panel of monoclonal anti-p53 antibodies showed that L12-derived clones that were transfected with the genomic p53 clone containe

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3

Dunn, Stephen J., Richard D. Oberst, Jeffrey L. Stott, and Bennie I. Osburn. "Molecular cloning of serogroup- and serotype-specific genome segments from bluetongue virus serotype 11." American Journal of Veterinary Research 50, no. 10 (1989): 1684–89. https://doi.org/10.2460/ajvr.1989.50.10.1684.

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SUMMARY Genome segments 2, 6, 8, and 9 of bluetongue virus (btv) serotype 11, coding for P2, NS1, NS2, and P6, respectively, were cloned into pUC 8. Sizes of segment-2 and segment-6 clones indicated partial copies (55% and 80% of full length, respectively), whereas segment 8 and 9 clones represented full-length copies. Northern blot hybridizations of the clones to the 5 United States btv prototypic serotypes (2, 10, 11, 13, and 17) revealed segment-2 clone to be serotype-specific to btv-11, whereas segment 6, 8, and 9 clones were able to detect all serotypes to varying degrees. All clones fail

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4

Nielsen, H. S., G. Liu, J. Nielsen, et al. "Generation of an Infectious Clone of VR-2332, a Highly Virulent North American-Type Isolate of Porcine Reproductive and Respiratory Syndrome Virus." Journal of Virology 77, no. 6 (2003): 3702–11. http://dx.doi.org/10.1128/jvi.77.6.3702-3711.2003.

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ABSTRACT A full-length cDNA clone of the prototypical North American porcine reproductive and respiratory syndrome virus (PRRSV) isolate VR-2332 was assembled in the plasmid vector pOK12. To rescue infectious virus, capped RNA was transcribed in vitro from the pOK12 clone and transfected into BHK-21C cells. The supernatant from transfected monolayers were serially passaged on Marc-145 cells and porcine pulmonary alveolar macrophages. Infectious PRRSV was recovered on Marc-145 cells as well as porcine pulmonary macrophages; thus, the cloned virus exhibited the same cell tropism as the parental

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5

Meulenberg, J. J. M., J. N. A. Bos-de Ruijter, R. van de Graaf, G. Wensvoort, and R. J. M. Moormann. "Infectious Transcripts from Cloned Genome-Length cDNA of Porcine Reproductive and Respiratory Syndrome Virus." Journal of Virology 72, no. 1 (1998): 380–87. http://dx.doi.org/10.1128/jvi.72.1.380-387.1998.

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ABSTRACT The 5′-terminal end of the genomic RNA of the Lelystad virus isolate (LV) of porcine reproductive and respiratory syndrome virus was determined. To construct full-length cDNA clones, the 5′-terminal sequence was ligated to cDNA clones covering the complete genome of LV. When RNA that was transcribed in vitro from these full-length cDNA clones was transfected into BHK-21 cells, infectious LV was produced and secreted. The virus was rescued by passage to porcine alveolar lung macrophages or CL2621 cells. When infectious transcripts were transfected to porcine alveolar lung macrophages o

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6

Liu, Guangqing, Yuying Zhang, Zheng Ni, et al. "Recovery of Infectious Rabbit Hemorrhagic Disease Virus from Rabbits after Direct Inoculation with In Vitro-Transcribed RNA." Journal of Virology 80, no. 13 (2006): 6597–602. http://dx.doi.org/10.1128/jvi.02078-05.

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ABSTRACT We report the first full-length infectious clone of strain JX/CHA/97 of rabbit hemorrhagic disease virus (RHDV). The transcripts from the full-length cDNA clones were infectious when they were directly injected into rabbits. The sequence of the virus recovered from the rabbits was identical to that of the injected RNA transcripts. The cDNA clone was engineered to contain one silent nucleotide change to create an EcoRV site (A to T at nucleotide 2908). The genetic marker was retained in the recovered progeny virus. The transfection of RNA transcripts into RK-13 cells resulted in the sy

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7

Wolf, D., Z. Laver-Rudich, and V. Rotter. "In vitro expression of human p53 cDNA clones and characterization of the cloned human p53 gene." Molecular and Cellular Biology 5, no. 8 (1985): 1887–93. http://dx.doi.org/10.1128/mcb.5.8.1887-1893.1985.

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The human p53 gene was cloned and characterized by using a battery of p53 DNA clones. A series of human cDNA clones of various sizes and relative localizations to the mRNA molecule were isolated by using the human p53-H14 (2.35-kilobase) cDNA probe which we previously cloned. One such isolate, clone p53-H7 (2.65 kilobases), spans the entire human mature p53 mRNA molecule. Construction of the human cDNA clones in the pSP65 RNA transcription vector facilitated the generation of p53 transcripts by the SP6 bacteriophage RNA polymerase. The p53-specific RNA transcripts obtained without further proc

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8

Wolf, D., Z. Laver-Rudich, and V. Rotter. "In vitro expression of human p53 cDNA clones and characterization of the cloned human p53 gene." Molecular and Cellular Biology 5, no. 8 (1985): 1887–93. http://dx.doi.org/10.1128/mcb.5.8.1887.

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The human p53 gene was cloned and characterized by using a battery of p53 DNA clones. A series of human cDNA clones of various sizes and relative localizations to the mRNA molecule were isolated by using the human p53-H14 (2.35-kilobase) cDNA probe which we previously cloned. One such isolate, clone p53-H7 (2.65 kilobases), spans the entire human mature p53 mRNA molecule. Construction of the human cDNA clones in the pSP65 RNA transcription vector facilitated the generation of p53 transcripts by the SP6 bacteriophage RNA polymerase. The p53-specific RNA transcripts obtained without further proc

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9

Lanford, Robert E., Helen Lee, Deborah Chavez, Bernadette Guerra, and Kathleen M. Brasky. "Infectious cDNA clone of the hepatitis C virus genotype 1 prototype sequence." Journal of General Virology 82, no. 6 (2001): 1291–97. http://dx.doi.org/10.1099/0022-1317-82-6-1291.

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A full-length cDNA clone of the hepatitis C virus (HCV) genotype 1 prototype (subtype 1a) sequence was constructed. Synthetic RNA produced from the initial cDNA clone was not infectious following intrahepatic inoculation of a chimpanzee. A consensus clone was prepared by comparison with multiple full-length HCV sequences of genotypes 1, 2 and 3. A total of 11 non-consensus amino acid residues were altered by mutagenesis. Synthetic RNA from the repaired clone initiated a typical, acute-resolving HCV infection following intrahepatic inoculation of a chimpanzee. In addition, at least one of three

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10

Harlow, E., N. M. Williamson, R. Ralston, D. M. Helfman, and T. E. Adams. "Molecular cloning and in vitro expression of a cDNA clone for human cellular tumor antigen p53." Molecular and Cellular Biology 5, no. 7 (1985): 1601–10. http://dx.doi.org/10.1128/mcb.5.7.1601-1610.1985.

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Three clones for the human tumor antigen p53 were isolated from a cDNA library prepared from A431 cells. One of these clones, pR4-2, contains the entire coding region for human p53. This clone directs the synthesis of a polypeptide with the correct molecular weight and immunological epitopes of an authentic p53 molecule in an in vitro transcription-translation reaction. Although the pR4-2 clone contains the coding region for p53, it is not a full-length copy of the human p53 mRNA. Northern analysis showed that the p53 mRNA is approximately 2,500 nucleotides long, whereas the pR4-2 insert is on

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11

Harlow, E., N. M. Williamson, R. Ralston, D. M. Helfman, and T. E. Adams. "Molecular cloning and in vitro expression of a cDNA clone for human cellular tumor antigen p53." Molecular and Cellular Biology 5, no. 7 (1985): 1601–10. http://dx.doi.org/10.1128/mcb.5.7.1601.

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Three clones for the human tumor antigen p53 were isolated from a cDNA library prepared from A431 cells. One of these clones, pR4-2, contains the entire coding region for human p53. This clone directs the synthesis of a polypeptide with the correct molecular weight and immunological epitopes of an authentic p53 molecule in an in vitro transcription-translation reaction. Although the pR4-2 clone contains the coding region for p53, it is not a full-length copy of the human p53 mRNA. Northern analysis showed that the p53 mRNA is approximately 2,500 nucleotides long, whereas the pR4-2 insert is on

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12

Shi, Pei-Yong, Mark Tilgner, Michael K. Lo, Kim A. Kent, and Kristen A. Bernard. "Infectious cDNA Clone of the Epidemic West Nile Virus from New York City." Journal of Virology 76, no. 12 (2002): 5847–56. http://dx.doi.org/10.1128/jvi.76.12.5847-5856.2002.

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ABSTRACT We report the first full-length infectious clone of the current epidemic, lineage I, strain of West Nile virus (WNV). The full-length cDNA was constructed from reverse transcription-PCR products of viral RNA from an isolate collected during the year 2000 outbreak in New York City. It was cloned into plasmid pBR322 under the control of a T7 promoter and stably amplified in Escherichia coli HB101. RNA transcribed from the full-length cDNA clone was highly infectious upon transfection into BHK-21 cells, resulting in progeny virus with titers of 1 × 109 to 5 × 109 PFU/ml. The cDNA clone w

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13

Chevalier, Sébastien Alain, Marine Walic, Sara Calattini, et al. "Construction and Characterization of a Full-Length Infectious Simian T-Cell Lymphotropic Virus Type 3 Molecular Clone." Journal of Virology 81, no. 12 (2007): 6276–85. http://dx.doi.org/10.1128/jvi.02538-06.

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ABSTRACT Together with their simian T-cell lymphotropic virus (STLV) equivalent, human T-cell lymphotropic virus type 1 (HTLV-1), HTLV-2, and HTLV-3 form the primate T-cell lymphotropic virus (PTLV) group. Over the years, understanding the biology and pathogenesis of HTLV-1 and HTLV-2 has been widely improved by the creation of molecular clones. In contrast, so far, PTLV-3 experimental studies have been restricted to the overexpression of the tax gene using reporter assays. We have therefore decided to construct an STLV-3 molecular clone. We generated a full-length STLV-3 proviral clone (8,891

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14

SANTOS, JEFFERSON J. S., MARLI T. CORDEIRO, GIOVANI R. BERTANI, ERNESTO T. A. MARQUES, and LAURA H. V. G. GIL. "A two-plasmid strategy for engineering a dengue virus type 3 infectious clone from primary Brazilian isolate." Anais da Academia Brasileira de Ciências 86, no. 4 (2014): 1749–59. http://dx.doi.org/10.1590/0001-3765201420130332.

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Dengue infections represent one of the most prevalent arthropod-borne diseases worldwide, causing a wide spectrum of clinical outcomes. Engineered infectious clone is an important tool to study Dengue virus (DENV) biology. Functional full-length cDNA clones have been constructed for many positive-strand RNA viruses and have provided valuable tools for studying the molecular mechanisms involved in viral genome replication, virion assembly, virus pathogenesis and vaccine development. We report herein the successful development of an infectious clone from a primary Brazilian isolate of dengue vir

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15

Tu, Liqin, Shuhua Wu, Danna Gao, Yong Liu, Yuelin Zhu, and Yinghua Ji. "Synthesis and Characterization of a Full-Length Infectious cDNA Clone of Tomato Mottle Mosaic Virus." Viruses 13, no. 6 (2021): 1050. http://dx.doi.org/10.3390/v13061050.

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Tomato mottle mosaic virus (ToMMV) is a noteworthy virus which belongs to the Virgaviridae family and causes serious economic losses in tomato. Here, we isolated and cloned the full-length genome of a ToMMV Chinese isolate (ToMMV-LN) from a naturally infected tomato (Solanum lycopersicum L.). Sequence analysis showed that ToMMV-LN contains 6399 nucleotides (nts) and is most closely related to a ToMMV Mexican isolate with a sequence identity of 99.48%. Next, an infectious cDNA clone of ToMMV was constructed by a homologous recombination approach. Both the model host N. benthamiana and the natur

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16

Nakada, T., I. Nagano, Z. Wu, and Y. Takahashi. "Molecular cloning and expression of the full-length tropomyosin gene from Trichinella spiralis." Journal of Helminthology 77, no. 1 (2003): 57–63. http://dx.doi.org/10.1079/joh2002153.

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AbstractA clone, designated as TsTM, was selected from the cDNA library of newborn larvae (NBL) of Trichinella spiralis through immunoscreening against infected sera. The clone contained a cDNA transcript of 855 bp in length with a single open reading frame, which encoded 285-amino acids (33 kDa in the estimated molecular weight). A sequence analysis revealed that the clone TsTM encoded the full-length of tropomyosin gene. The phylogenetic analysis of the tropomyosin gene was in good agreement with the classical taxonomical position of T. spiralis. The fusion proteins encoded by the clone TsTM

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17

Elborough, K. M., R. Swinhoe, R. Winz, et al. "Isolation of cDNAs from Brassica napus encoding the biotin-binding and transcarboxylase domains of acetyl-CoA carboxylase: assignment of the domain structure in a full-length Arabidopsis thaliana genomic clone." Biochemical Journal 301, no. 2 (1994): 599–605. http://dx.doi.org/10.1042/bj3010599.

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One independent and two overlapping rape cDNA clones have been isolated from a rape embryo library. We have shown that they encode a 2.3 kb and a 2.5 kb stretch of the full-length acetyl-CoA carboxylase (ACCase) cDNA, corresponding to the biotin-binding and transcarboxylase domains respectively. Using the cDNA in Northern-blot analysis we have shown that the mRNA for ACCase has a higher level of expression in rape seed than in rape leaf and has a full length of 7.5 kb. The level of expression during rape embryogenesis was compared with both oil deposition and expression of two fatty acid synth

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18

Poirier, John T., P. Seshidhar Reddy, Neeraja Idamakanti, et al. "Characterization of a full-length infectious cDNA clone and a GFP reporter derivative of the oncolytic picornavirus SVV-001." Journal of General Virology 93, no. 12 (2012): 2606–13. http://dx.doi.org/10.1099/vir.0.046011-0.

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Seneca Valley virus (SVV-001) is an oncolytic picornavirus with selective tropism for a subset of human cancers with neuroendocrine differentiation. To characterize further the specificity of SVV-001 and its patterns and kinetics of intratumoral spread, bacterial plasmids encoding a cDNA clone of the full-length wild-type virus and a derivative virus expressing GFP were generated. The full-length cDNA of the SVV-001 RNA genome was cloned into a bacterial plasmid under the control of the T7 core promoter sequence to create an infectious cDNA clone, pNTX-09. A GFP reporter virus cDNA clone, pNTX

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19

Haviland, D. L., J. C. Haviland, D. T. Fleischer, A. Hunt, and R. A. Wetsel. "Complete cDNA sequence of human complement pro-C5. Evidence of truncated transcripts derived from a single copy gene." Journal of Immunology 146, no. 1 (1991): 362–68. http://dx.doi.org/10.4049/jimmunol.146.1.362.

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Abstract Two truncated human C5 clones, pHC5A and pHC5B, were isolated from an adult human liver cDNA library, and contained inserts of 2930 and 2181 bp, respectively. Both clones were polyadenylated and encoded the 5'-end of the C5 pro-molecule, thereby completing the human pro-C5 cDNA sequence. However, near the 3'-ends, at exon/intron boundaries, the nucleotide sequences of pHC5A and pHC5B diverged from each other and from the full-length 6.0-kb C5 cDNA sequence. Clone pHC5A, which overlapped the first human C5 clone described (J-16), encoded most of the C5 signal peptide, the complete beta

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20

Maughan, Peter J., Scott M. Smith, and Joshua A. Raney. "Utilization of Super BAC Pools and Fluidigm Access Array Platform for High-Throughput BAC Clone Identification: Proof of Concept." Journal of Biomedicine and Biotechnology 2012 (2012): 1–7. http://dx.doi.org/10.1155/2012/405940.

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Bacterial artificial chromosome (BAC) libraries are critical for identifying full-length genomic sequences, correlating genetic and physical maps, and comparative genomics. Here we describe the utilization of the Fluidigm access array genotyping system in conjunction with KASPar genotyping technology to identify individual BAC clones corresponding to specific single-nucleotide polymorphisms (SNPs) from an Amplicon Express seven-plate super pooledAmaranthus hypochondriacusBAC library. Ninety-six SNP loci, spanning the length ofA. hypochondriacuslinkage groups 1, 2, and 15, were simultaneously t

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Cordes, K., E. Maiss, S. Winter, and H. Rose. "Complete genome sequence and construction of an infectious full-length cDNA clone of a cucumber vein yellowing virus (CVYV) isolate from Portugal." Archives of Virology 166, no. 12 (2021): 3417–20. http://dx.doi.org/10.1007/s00705-021-05248-y.

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AbstractCucumber vein yellowing virus (CVYV) is a member of the genus Ipomovirus in the family Potyviridae. In the National Center for Biotechnology Information (NCBI) database, three complete genome sequences of CVYV isolates from Spain (NC_006941), Israel (KT276369), and Jordan (JF460793) are available. In this study, we report the complete sequence of an isolate of CVYV from Portugal (DSMZ PV-0776) along with the construction of an infectious full-length cDNA clone via Gibson assembly. The sequence of CVYV Portugal shows the closest relationship to a CVYV isolate from Spain (genome, 99.7% i

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Cordes, K., E. Maiss, S. Winter, and H. Rose. "Complete genome sequence and construction of an infectious full-length cDNA clone of a cucumber vein yellowing virus (CVYV) isolate from Portugal." Archives of Virology 166, no. 12 (2021): 3417–20. http://dx.doi.org/10.1007/s00705-021-05248-y.

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AbstractCucumber vein yellowing virus (CVYV) is a member of the genus Ipomovirus in the family Potyviridae. In the National Center for Biotechnology Information (NCBI) database, three complete genome sequences of CVYV isolates from Spain (NC_006941), Israel (KT276369), and Jordan (JF460793) are available. In this study, we report the complete sequence of an isolate of CVYV from Portugal (DSMZ PV-0776) along with the construction of an infectious full-length cDNA clone via Gibson assembly. The sequence of CVYV Portugal shows the closest relationship to a CVYV isolate from Spain (genome, 99.7% i

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23

Kunapuli, S. P., G. Fen Mao, M. Bastepe, et al. "Cloning and expression of a prostaglandin E receptor EP3 subtype from human erythroleukaemia cells." Biochemical Journal 298, no. 2 (1994): 263–67. http://dx.doi.org/10.1042/bj2980263.

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Prostaglandins inhibit platelet activation by stimulating intracellular cyclic AMP formation. We have postulated that intracellular cyclic AMP levels in platelets are buffered by a distinct prostaglandin receptor that mediates inhibition of cyclic AMP formation. In order to provide evidence for the model, we have cloned the cDNA coding for a prostaglandin receptor EP3 subtype, which is coupled to inhibition of adenylate cyclase, from the megakaryocytic cell line human erythroleukaemia (HEL) cells. A PCR-generated hybridization probe, produced using primers based on the sequence of the mouse pr

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Sbardellati, Andrea, Elisa Scarselli, Ernst Verschoor, Amedeo De Tomassi, Domenico Lazzaro, and Cinzia Traboni. "Generation of infectious and transmissible virions from a GB virus B full-length consensus clone in tamarins." Journal of General Virology 82, no. 10 (2001): 2437–48. http://dx.doi.org/10.1099/0022-1317-82-10-2437.

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The strong similarity between GB virus B (GBV-B) and hepatitis C virus (HCV) makes tamarins infected by GBV-B an acceptable surrogate animal model for HCV infection. Even more attractive, for drug discovery purposes, is the idea of constructing chimeric viruses by inserting HCV genes of interest into a GBV-B genome frame. To accomplish this, infectious cDNA clones of both viruses must be available. The characterization of several HCV molecular clones capable of infecting chimpanzees has been published, whereas only one infectious GBV-B clone inducing hepatitis in tamarins has been reported so

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Voliva, C. F., and K. Paigen. "Isolation of the mouse cytochrome P450J (CYP2E1) cDNA and its reciprocal testosterone regulation in kidney and liver." Journal of Molecular Endocrinology 7, no. 2 (1991): 155–66. http://dx.doi.org/10.1677/jme.0.0070155.

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ABSTRACT A hybridization probe that is homologous to the B2 short interspersed repetitive element detects an mRNA in mouse kidney and liver that is regulated by androgen. Administration of testosterone induces this mRNA in kidney and represses it in liver. The mRNA was cloned by first using the B2 probe to select 48 cDNA clones from an androgen-induced kidney library. These clones were then tested for their androgen response by hybridizing them with probes made by reverse transcription of basal and testosterone-treated kidney poly(A)+ RNA. Any homology to the B2 sequence was masked by prehybri

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Zhang, Cheng, Fanlei Ran, Lei Du, et al. "The Humanization and Maturation of an Anti-PrPc Antibody." Bioengineering 11, no. 3 (2024): 242. http://dx.doi.org/10.3390/bioengineering11030242.

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The cellular prion protein (PrPc) is a cell surface glycoprotein that is highly expressed in a variety of cancer tissues in addition to the nervous system, and its elevated expression is correlated to poor prognosis in many cancer patients. Our team previously found that patients with colorectal cancer (CRC) with high-level PrPc expression had significantly poorer survival than those with no or low-level PrPc expression. Mouse antibodies for PrPc inhibited tumor initiation and liver metastasis of PrPc-positive human CRC cells in mouse model experiments. PrPc is a candidate target for CRC thera

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Shirasaki, Ryosuke, Haruko Tashiro, Mitsuho Noguchi, Moritaka Goto, Kazuo Kawasugi, and Naoki Shirafuji. "Isolation and Characterization of Murine Novel cDNA Clone Using Newly Constructed Gene Trap Vector." Blood 106, no. 11 (2005): 4229. http://dx.doi.org/10.1182/blood.v106.11.4229.4229.

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Abstract Objective: To understand better the mechanism of the maintenance of immature state of stem cells, murine novel cDNA clones are to be isolated by the gene trap method. Method: The trap vector was constructed with En-2 intron sequence followed by splicing acceptor sequence, IRES signal sequence and b-galactosidase cDNA. NeoR gene was also contained without poly-A additional signal. After electroporation into Embryonic stem (Es) cell lines (D3, and E14.1 Köln), neomycin-resistant Es clones were picked up, and were selected in which the expression of b-gal was detected in the undifferent

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28

Sun, Yuning, Aaron Yun Chen, Fang Cheng, Wuxiang Guan, F. Brent Johnson, and Jianming Qiu. "Molecular Characterization of Infectious Clones of the Minute Virus of Canines Reveals Unique Features of Bocaviruses." Journal of Virology 83, no. 8 (2009): 3956–67. http://dx.doi.org/10.1128/jvi.02569-08.

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ABSTRACT Minute virus of canines (MVC) is a member of the genus Bocavirus in the family Parvoviridae. We have molecularly cloned and sequenced the 5′- and 3′-terminal palindromes of MVC. The MVC genome, 5,404 nucleotides (nt) in length, shared an identity of 52.6% and 52.1% with that of human bocavirus and bovine parvovirus, respectively. It had distinct palindromic hairpins of 183 nt and 198 nt at the left-end and right-end termini of the genome, respectively. The left-end terminus was also found in two alternative orientations (flip or flop). Both termini shared extensive similarities with t

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Yount, Boyd, Kristopher M. Curtis, and Ralph S. Baric. "Strategy for Systematic Assembly of Large RNA and DNA Genomes: Transmissible Gastroenteritis Virus Model." Journal of Virology 74, no. 22 (2000): 10600–10611. http://dx.doi.org/10.1128/jvi.74.22.10600-10611.2000.

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ABSTRACT A systematic method was developed to assemble functional full-length genomes of large RNA and DNA viruses. Coronaviruses contain the largest single-stranded positive-polarity RNA genome in nature. The ∼30-kb genome, coupled with regions of genomic instability, has hindered the development of a full-length infectious cDNA construct. We have assembled a full-length infectious construct of transmissible gastroenteritis virus (TGEV), an important pathogen in swine. Using a novel approach, six adjoining cDNA subclones that span the entire TGEV genome were isolated. Each clone was engineere

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30

Joswig, G., C. Petzelt, and D. Werner. "Murine cDNAs coding for the centrosomal antigen centrosomin A." Journal of Cell Science 98, no. 1 (1991): 37–43. http://dx.doi.org/10.1242/jcs.98.1.37.

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Screening of an induced Ehrlich ascites cell-derived lambda gt11 cDNA library with an antibody (GP1), immunoreacting specifically with centrosomal antigen(s) of interphase and mitotic cells of different species, released a partial cDNA clone (lambda P10A) encoding the carboxy-terminal section of a centrosome-specific antigen. This specificity of the clone lambda P10A could be verified by lacZ-directed antigen expression from Escherichia coli Y1089 lysogenized with the recombinant phage lambda P10A and subsequent production of centrosome-specific antibodies by means of the recombinant antigen.

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Huang, Y. W., G. Haqshenas, C. Kasorndorkbua, P. G. Halbur, S. U. Emerson, and X. J. Meng. "Capped RNA Transcripts of Full-Length cDNA Clones of Swine Hepatitis E Virus Are Replication Competent When Transfected into Huh7 Cells and Infectious When Intrahepatically Inoculated into Pigs." Journal of Virology 79, no. 3 (2005): 1552–58. http://dx.doi.org/10.1128/jvi.79.3.1552-1558.2005.

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ABSTRACT Swine hepatitis E virus (swine HEV), the first animal strain of HEV to be isolated, is a zoonotic agent. We report here the construction and in vitro and in vivo characterizations of infectious cDNA clones of swine HEV. Eight overlapping fragments spanning the entire genome were amplified by reverse transcription-PCR and assembled into a full-length cDNA clone, clone C, which contained 14 mutations compared to the consensus sequence of swine HEV. RNA transcripts from clone C were not infectious, as determined by intrahepatic inoculation into pigs and by in vitro transfection of Huh7 c

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32

de Wet, J. R., K. V. Wood, M. DeLuca, D. R. Helinski, and S. Subramani. "Firefly luciferase gene: structure and expression in mammalian cells." Molecular and Cellular Biology 7, no. 2 (1987): 725–37. http://dx.doi.org/10.1128/mcb.7.2.725-737.1987.

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The nucleotide sequence of the luciferase gene from the firefly Photinus pyralis was determined from the analysis of cDNA and genomic clones. The gene contains six introns, all less than 60 bases in length. The 5' end of the luciferase mRNA was determined by both S1 nuclease analysis and primer extension. Although the luciferase cDNA clone lacked the six N-terminal codons of the open reading frame, we were able to reconstruct the equivalent of a full-length cDNA using the genomic clone as a source of the missing 5' sequence. The full-length, intronless luciferase gene was inserted into mammali

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de Wet, J. R., K. V. Wood, M. DeLuca, D. R. Helinski, and S. Subramani. "Firefly luciferase gene: structure and expression in mammalian cells." Molecular and Cellular Biology 7, no. 2 (1987): 725–37. http://dx.doi.org/10.1128/mcb.7.2.725.

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The nucleotide sequence of the luciferase gene from the firefly Photinus pyralis was determined from the analysis of cDNA and genomic clones. The gene contains six introns, all less than 60 bases in length. The 5' end of the luciferase mRNA was determined by both S1 nuclease analysis and primer extension. Although the luciferase cDNA clone lacked the six N-terminal codons of the open reading frame, we were able to reconstruct the equivalent of a full-length cDNA using the genomic clone as a source of the missing 5' sequence. The full-length, intronless luciferase gene was inserted into mammali

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Donaldson, Eric F., Boyd Yount, Amy C. Sims, Susan Burkett, Raymond J. Pickles, and Ralph S. Baric. "Systematic Assembly of a Full-Length Infectious Clone of Human Coronavirus NL63." Journal of Virology 82, no. 23 (2008): 11948–57. http://dx.doi.org/10.1128/jvi.01804-08.

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ABSTRACT Historically, coronaviruses were predominantly associated with mild upper respiratory disease in humans. More recently, three novel coronaviruses associated with severe human respiratory disease were found, including (i) the severe acute respiratory syndrome coronavirus, associated with a significant atypical pneumonia and 10% mortality; (ii) HKU-1, associated with chronic pulmonary disease; and (iii) NL63, associated with both upper and lower respiratory tract disease in children and adults worldwide. These discoveries establish coronaviruses as important human pathogens and undersco

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Day, A. J., J. Ripoche, A. Lyons, B. McIntosh, T. J. R. Harris, and R. B. Sim. "Sequence analysis of a cDNA clone encoding the C-terminal end of human complement factor H." Bioscience Reports 7, no. 3 (1987): 201–7. http://dx.doi.org/10.1007/bf01124790.

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Peptide sequencing of the complement system regulatory protein, factor H, permitted the synthesis of a mixed sequence oligonucleotide probe. Human liver cDNA libraries were screened and factor H-specific clones selected. No full-length clone was obtained, but the largest available clone, R2a, was found to encode the C-terminal 657 amino acids of factor H. The derived amino acid sequence consists of 10 contiguous internally homologous segments, each about 60 amino acids long. Sequences homologous to these are found in several other complement and non-complement proteins. Such sequences are like

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Tabler, Martin, Martina Schnölzer, and Heinz L. Sänger. "Molecular cloning of potato spindle tuber viroid (PSTV) cDNA synthesized by enzymatic elongation of PSTV-specific DNA primers: A general strategy for viroid cloning." Bioscience Reports 5, no. 2 (1985): 143–58. http://dx.doi.org/10.1007/bf01117061.

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Different cDNAs were synthesized by primer extension from the RNA of the severe strain KF 440 of potato spindle tuber viroid (PSTV) with the aid of reverse transcriptase using three PSTV-specific DNA molecules as primers. The cDNAs were made double-stranded and cloned into plasmid pBR 322. Various overlapping subgenomic DNA fragments were prepared from these clones and recombined in two different ways. In both cases a PSTV DNA copy was obtained which represented the entire PSTV RNA genome. The sequence of the DNA of one of the resulting full-length clones was identical with the original PSTV i

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Franck, Jens P. C., Jeffery Morrissette, John E. Keen, Richard L. Londraville, Mark Beamsley, and Barbara A. Block. "Cloning and characterization of fiber type-specific ryanodine receptor isoforms in skeletal muscles of fish." American Journal of Physiology-Cell Physiology 275, no. 2 (1998): C401—C415. http://dx.doi.org/10.1152/ajpcell.1998.275.2.c401.

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We have cloned a group of cDNAs that encodes the skeletal ryanodine receptor isoform (RyR1) of fish from a blue marlin extraocular muscle library. The cDNAs encode a protein of 5,081 amino acids with a calculated molecular mass of 576,302 Da. The deduced amino acid sequence shows strong sequence identity to previously characterized RyR1 isoforms. An RNA probe derived from a clone of the full-length marlin RyR1 isoform hybridizes to RNA preparations from extraocular muscle and slow-twitch skeletal muscle but not to RNA preparations from fast-twitch skeletal or cardiac muscle. We have also isola

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Kaneda, Norio, Hiroshi Ichinose, Kazuto Kobayashi, et al. "Isolation of a full-length cDNA clone encoding human phenylethanolamine -methyltransferase." Neuroscience Research Supplements 7 (January 1988): S116. http://dx.doi.org/10.1016/0921-8696(88)90242-3.

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Arai, N., D. Nomura, K. Yokota, et al. "Immunologically distinct p53 molecules generated by alternative splicing." Molecular and Cellular Biology 6, no. 9 (1986): 3232–39. http://dx.doi.org/10.1128/mcb.6.9.3232-3239.1986.

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Transfection of a functional cloned p53 gene into an L12 p53 nonproducer cell line efficiently reconstituted p53 expression. The p53 protein synthesized in these clones was indistinguishable from that occurring naturally in tumor cells. When a p53 cDNA clone was used instead, we observed that the L12-derived clones exhibited a distinct immunological profile. In the present experiments we compared the immunological epitopes of p53 proteins encoded by several full-length cDNA clones. Immunoprecipitation of p53 proteins generated by in vitro transcription and translation of the various cDNA clone

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Arai, N., D. Nomura, K. Yokota, et al. "Immunologically distinct p53 molecules generated by alternative splicing." Molecular and Cellular Biology 6, no. 9 (1986): 3232–39. http://dx.doi.org/10.1128/mcb.6.9.3232.

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Transfection of a functional cloned p53 gene into an L12 p53 nonproducer cell line efficiently reconstituted p53 expression. The p53 protein synthesized in these clones was indistinguishable from that occurring naturally in tumor cells. When a p53 cDNA clone was used instead, we observed that the L12-derived clones exhibited a distinct immunological profile. In the present experiments we compared the immunological epitopes of p53 proteins encoded by several full-length cDNA clones. Immunoprecipitation of p53 proteins generated by in vitro transcription and translation of the various cDNA clone

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Wang, Yong, Zhenggang Wang, Juan Li, Yajun Wang, and Frederick C. C. Leung. "Database for chicken full-length cDNAs." Physiological Genomics 28, no. 2 (2007): 141–45. http://dx.doi.org/10.1152/physiolgenomics.00097.2006.

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The generation of full-length cDNA databases is essential for functional genomics studies as well as for correct annotation of species genomic sequences. Human and mouse full-length cDNA projects have provided the biomedical research community with a large amount of gene information. Recent completion of the chicken genome sequence draft now enables a similar full-length cDNA project to be initiated for this species. In this report, we introduce the development of a chicken full-length cDNA database, which will facilitate future research work in this biological system. In this project, chicken

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Córdoba, Laura, Alicia R. Feagins, Tanja Opriessnig, et al. "Rescue of a genotype 4 human hepatitis E virus from cloned cDNA and characterization of intergenotypic chimeric viruses in cultured human liver cells and in pigs." Journal of General Virology 93, no. 10 (2012): 2183–94. http://dx.doi.org/10.1099/vir.0.043711-0.

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Hepatitis E virus (HEV) is an important but extremely understudied human pathogen. Genotypes 1 and 2 are restricted to humans, whereas genotypes 3 and 4 are zoonotic, infecting both humans and pigs. This report describes, for the first time, the successful rescue of infectious HEV in vitro and in vivo from cloned cDNA of a genotype 4 human HEV (strain TW6196E). The complete genomic sequence of the TW6196E virus was determined and a full-length cDNA clone (pHEV-4TW) was assembled. Capped RNA transcripts from the pHEV-4TW clone were replication competent in Huh7 cells and infectious in HepG2/C3A

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Mizuno, S., J. A. Trapani, B. H. Koller, B. Dupont, and S. Y. Yang. "Isolation and nucleotide sequence of a cDNA clone encoding a novel HLA class I gene." Journal of Immunology 140, no. 11 (1988): 4024–30. http://dx.doi.org/10.4049/jimmunol.140.11.4024.

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Abstract A cDNA clone, designated JTW15, has been isolated and found to encode a novel, divergent class I HLA gene transcript. The clone was identified by its ability to hybridize with a pan-HLA class I nucleotide probe while failing to bind probes specific for the 3' untranslated exons of the HLA-A, -B, and -C genes. JTW15, 1665 nucleotides in length, represents a full length copy of an approximately 1.7-kb mRNA detected in Northern blot analyses. Comparison of the derived amino acid sequence with those of other HLA class I proteins showed JTW15 to be the most divergent HLA class I gene ident

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McElroy, Kate L., Konstantin A. Tsetsarkin, Dana L. Vanlandingham, and Stephen Higgs. "Characterization of an infectious clone of the wild-type yellow fever virus Asibi strain that is able to infect and disseminate in mosquitoes." Journal of General Virology 86, no. 6 (2005): 1747–51. http://dx.doi.org/10.1099/vir.0.80746-0.

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Infectious clone technology provides an opportunity to study the molecular basis of arthropod–virus interactions in detail. This study describes the development of an infectious clone of the prototype yellow fever virus Asibi strain (YFV-As) with the purpose of identifying sequences or domains that influence infection dynamics in the mosquito vector. The full-length cDNA of YFV-As virus was produced from RT-PCR products of parental viral RNA. These were cloned into a low-copy-number plasmid previously used to develop the YFV-17D infectious clone (pACNR/FLYF-17D). Virus recovered from the infec

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Simkovich, Aaron J., Yinzi Li, Susanne E. Kohalmi, Jonathan S. Griffiths, and Aiming Wang. "Molecular Identification of Prune Dwarf Virus (PDV) Infecting Sweet Cherry in Canada and Development of a PDV Full-Length Infectious cDNA Clone." Viruses 13, no. 10 (2021): 2025. http://dx.doi.org/10.3390/v13102025.

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Prune dwarf virus (PDV) is a member of ilarviruses that infects stone fruit species such as cherry, plum and peach, and ornamentally grown trees worldwide. The virus lacks an RNA silencing suppressor. Infection by PDV either alone, or its mixed infection with other viruses causes deteriorated fruit marketability and reduced fruit yields. Here, we report the molecular identification of PDV from sweet cherry in the prominent fruit growing region of Ontario, Canada known as the Niagara fruit belt using next generation sequencing of small interfering RNAs (siRNAs). We assessed its incidence in an

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Baharudeen, Zamrina, Rahmah Noordin, Lim Theam Soon, et al. "Isolation and Production of Human Monoclonal Antibody Proteins against a Toxocara canis Excretory–Secretory Recombinant Antigen." Pathogens 11, no. 11 (2022): 1232. http://dx.doi.org/10.3390/pathogens11111232.

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Toxocariasis is a widespread zoonotic parasitic disease with a significant socioeconomic impact, particularly on underprivileged communities. Limitations of existing diagnostic tools and vague presenting symptoms may lead to misdiagnosis, thus underestimating the actual global impact of the disease. The present study describes the isolation and production of novel recombinant monoclonal antibodies against Toxocara canis recombinant TES-26 antigen (rTES-26) utilizing a human helminth scFv phage display library. The isolated antibody clones were characterized based on their gene sequences and bi

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James, Anthony P., Dawit B. Kidanemariam, Sharon D. Hamill, James L. Dale, and Robert M. Harding. "Infectivity of an Infectious Clone of Banana Streak CA Virus in A-Genome Bananas (Musa acuminata ssp.)." Viruses 13, no. 6 (2021): 1071. http://dx.doi.org/10.3390/v13061071.

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We have characterized the complete genome sequence of an Australian isolate of banana streak CA virus (BSCAV). A greater-than-full-length, cloned copy of the virus genome was assembled and agroinoculated into five tissue-cultured plants of nine different Musa acuminata banana accessions. BSCAV was highly infectious in all nine accessions. All five inoculated plants from eight accessions developed symptoms by 28 weeks post-inoculation, while all five plants of M. acuminata AA subsp. zebrina remained symptomless. Symptoms were mild in six accessions but were severe in Khae Phrae (M. acuminata su

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Smeekens, Sjef, Jan van Binsbergen, and Peter Weisbeck. "The plant ferredoxin precursor: nucleotide sequence of a full length cDNA clone." Nucleic Acids Research 13, no. 9 (1985): 3179–94. http://dx.doi.org/10.1093/nar/13.9.3179.

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Himeda, Toshiki, Takushi Hosomi, Naeem Asif, et al. "The preparation of an infectious full-length cDNA clone of Saffold virus." Virology Journal 8, no. 1 (2011): 110. http://dx.doi.org/10.1186/1743-422x-8-110.

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Wetmur, J. G., D. F. Bishop, C. Cantelmo, and R. J. Desnick. "Human delta-aminolevulinate dehydratase: nucleotide sequence of a full-length cDNA clone." Proceedings of the National Academy of Sciences 83, no. 20 (1986): 7703–7. http://dx.doi.org/10.1073/pnas.83.20.7703.

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