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Journal articles on the topic 'FLAG epitope tag'

Author: Grafiati

Published: 4 June 2021

Last updated: 5 February 2022

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1

Guo, Xin-Yu, Xiao-Dong Gao, and Morihisa Fujita. "Sulfation of a FLAG tag mediated by SLC35B2 and TPST2 affects antibody recognition." PLOS ONE 16, no. 5 (2021): e0250805. http://dx.doi.org/10.1371/journal.pone.0250805.

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A FLAG tag consisting of DYKDDDDK is an epitope tag that is frequently and widely used to detect recombinant proteins of interest. In this study, we performed a CRISPR-based genetic screening to identify factors involved in the detection of a FLAG-tagged misfolded model protein at the cell surface. In the screening, SLC35B2, which encodes 3’-phosphoadenosine-5’-phosphosulfate transporter 1, was identified as the candidate gene. The detection of FLAG-tagged misfolded proteins at the cell surface was significantly increased in SLC35B2-knockout cells. Furthermore, protein tyrosine sulfation media

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2

Zhao, Xinyu, Guoshun Li, and Shufang Liang. "Several Affinity Tags Commonly Used in Chromatographic Purification." Journal of Analytical Methods in Chemistry 2013 (2013): 1–8. http://dx.doi.org/10.1155/2013/581093.

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Affinity tags have become powerful tools from basic biological research to structural and functional proteomics. They were widely used to facilitate the purification and detection of proteins of interest, as well as the separation of protein complexes. Here, we mainly discuss the benefits and drawbacks of several affinity or epitope tags frequently used, including hexahistidine tag, FLAG tag, Strep II tag, streptavidin-binding peptide (SBP) tag, calmodulin-binding peptide (CBP), glutathione S-transferase (GST), maltose-binding protein (MBP), S-tag, HA tag, and c-Myc tag. In some cases, a large

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Seago, Julian, Terry Jackson, Claudia Doel, et al. "Characterization of epitope-tagged foot-and-mouth disease virus." Journal of General Virology 93, no. 11 (2012): 2371–81. http://dx.doi.org/10.1099/vir.0.043521-0.

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Foot-and-mouth disease (FMD) is a highly contagious and economically devastating disease of cloven-hoofed animals with an almost-worldwide distribution. Conventional FMD vaccines consisting of chemically inactivated viruses have aided in the eradication of FMD from Europe and remain the main tool for control in endemic countries. Although significant steps have been made to improve the quality of vaccines, such as improved methods of antigen concentration and purification, manufacturing processes are technically demanding and expensive. Consequently, there is large variation in the quality of

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4

Laird, Melissa E., and Ronald C. Desrosiers. "Infectivity and Neutralization of Simian Immunodeficiency Virus with FLAG Epitope Insertion in gp120 Variable Loops." Journal of Virology 81, no. 20 (2007): 10838–48. http://dx.doi.org/10.1128/jvi.00831-07.

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ABSTRACT A FLAG epitope tag was substituted within variable loop 1 (V1), 2 (V2), or 4 (V4) of the gp120 envelope glycoprotein of simian immunodeficiency virus strain 239 (SIV239) to evaluate the extent to which each variable loop may serve as a target for antibody-mediated neutralization. Two sites within each variable loop of SIV239 were chosen for individual epitope tag insertions. FLAG epitope substitutions were also made in the V1, V2, and V4 loops of a neutralization-sensitive derivative of SIV239, SIV316. Of the 10 FLAG-tagged recombinant viruses analyzed, three (SIV239FV1b, SIV239FV2b,

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5

Plemper, Richard K., Anthea L. Hammond, Denis Gerlier, Adele K. Fielding, and Roberto Cattaneo. "Strength of Envelope Protein Interaction Modulates Cytopathicity of Measles Virus." Journal of Virology 76, no. 10 (2002): 5051–61. http://dx.doi.org/10.1128/jvi.76.10.5051-5061.2002.

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ABSTRACT To understand the molecular determinants of measles virus (MV) cytopathicity, we have characterized mutant viruses exhibiting a more-extensive cell-to-cell fusion while maintaining efficient replication to high titers. A virus which is modified by the addition of an 8-amino-acid Flag epitope tag at the cytoplasmic tail of its H (for MV hemagglutinin) envelope glycoprotein replicates efficiently, has an increased cytopathicity, possesses a greater infectivity per particle, and has an altered protein composition compared with that of unmodified MV. The mutant phenotype is not specifical

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6

BLONG, M. Renee, Elliott BEDOWS, and Oksana LOCKRIDGE. "Tetramerization domain of human butyrylcholinesterase is at the C-terminus." Biochemical Journal 327, no. 3 (1997): 747–57. http://dx.doi.org/10.1042/bj3270747.

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Butyrylcholinesterase (BChE) in human serum consists predominantly of tetramers. Recombinant BChE, however, expressed in Chinese hamster ovary (CHO) cells, consists of approx. 55% dimers, 10-30% tetramers and 15-40% monomers. To determine the origin of the monomer species we added the FLAG epitope (epitope tag, amino acid sequence DYKDDDDK) to the C-terminus of the enzyme, and expressed BChE-FLAG in CHO cells. We found that secreted, active monomers had lost their FLAG epitope, suggesting that the monomers were made by proteolysis of dimers or tetramers at the C-terminus. To estimate the numbe

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7

Joyce, Charles W., Gregory S. Shelness, Matthew A. Davis, et al. "ACAT1 and ACAT2 Membrane Topology Segregates a Serine Residue Essential for Activity to Opposite Sides of the Endoplasmic Reticulum Membrane." Molecular Biology of the Cell 11, no. 11 (2000): 3675–87. http://dx.doi.org/10.1091/mbc.11.11.3675.

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A second form of the enzyme acyl-CoA:cholesterol acyltransferase, ACAT2, has been identified. To explore the hypothesis that the two ACAT enzymes have separate functions, the membrane topologies of ACAT1 and ACAT2 were examined. A glycosylation reporter and FLAG epitope tag sequence was appended to a series of ACAT cDNAs truncated after each predicted transmembrane domain. Fusion constructs were assembled into microsomal membranes, in vitro, and topologies were determined based on glycosylation site use and accessibility to exogenous protease. The accessibility of the C-terminal FLAG epitope i

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8

Mazurov, Dmitriy, Alexandra Maslennikova, Dmitriy Komkov, and Anastasia Zotova. "Application of SORTS, a Novel Gene-Edited Cell Selection Method for HIV Study and Therapy." Proceedings 50, no. 1 (2020): 13. http://dx.doi.org/10.3390/proceedings2020050013.

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We have recently developed surface oligopeptide knock-in for rapid target selection (SORTS), a novel method to isolate mammalian cells with gene modifications using FACS-sorting. It relies on CRISPR/Cas9-mediated targeted knock-in of a very short promoterless expression construct (250 bp) comprising a Flag or HA epitope embedded into the smallest GPI-protein CD52, and a polyA signal from the beta-globin. CD52 efficiently delivers the epitope to the cell surface, where it serves as a marker for selection, while polyA terminates transcription and silences target gene expression. Primarily, SORTS

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9

Elvira, Bernat, Jamshed Warsi, Myriam Fezai, Carlos Munoz, and Florian Lang. "SPAK and OSR1 Sensitive Cell Membrane Protein Abundance and Activity of KCNQ1/E1 K+ Channels." Cellular Physiology and Biochemistry 37, no. 5 (2015): 2032–42. http://dx.doi.org/10.1159/000438563.

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Background/Aims: KCNQ1/E1 channels are expressed in diverse tissues and serve a variety of functions including endolymph secretion in the inner ear, cardiac repolarization, epithelial transport and cell volume regulation. Kinases involved in regulation of epithelial transport and cell volume include SPAK (SPS1-related proline/alanine-rich kinase) and OSR1 (oxidative stress-responsive kinase 1), which are under control of WNK (with-no-K[Lys]) kinases. The present study explored whether KCNQ1/E1 channels are regulated by SPAK and/or OSR1. Methods: cRNA encoding KCNQ1/E1 was injected into Xenopus

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10

Kaltwasser, Marcus, Thomas Wiegert, and Wolfgang Schumann. "Construction and Application of Epitope- and Green Fluorescent Protein-Tagging Integration Vectors for Bacillus subtilis." Applied and Environmental Microbiology 68, no. 5 (2002): 2624–28. http://dx.doi.org/10.1128/aem.68.5.2624-2628.2002.

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ABSTRACT Here we describe the construction and application of six new tagging vectors allowing the fusion of two different types of tagging sequences, epitope and localization tags, to any Bacillus subtilis protein. These vectors are based on the backbone of pMUTIN2 and replace the lacZ gene with tagging sequences. Fusion of the tagging sequences occurs by PCR amplification of the 3′ terminal part of the gene of interest (about 300 bp), insertion into the tagging vector in such a way that a fusion protein will be synthesized upon integration of the whole vector via homologous recombination wit

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11

Wegelt, Anne, Ilona Reimann, Harald Granzow, and Martin Beer. "Characterization and purification of recombinant bovine viral diarrhea virus particles with epitope-tagged envelope proteins." Journal of General Virology 92, no. 6 (2011): 1352–57. http://dx.doi.org/10.1099/vir.0.029330-0.

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Bovine viral diarrhea virus (BVDV) belongs to the genus Pestivirus within the family Flaviviridae. The lipid membrane of the virions is supposed to contain the three glycosylated envelope proteins Erns, E1 and E2, but detailed studies of virus assembly are complicated because no efficient purification method for pestiviruses has been described so far. In this study, we generated infectious BVDV with N-terminally FLAG-tagged Erns or E2 proteins, respectively. The expression of the epitope-tagged Erns and E2 proteins could be shown by immunofluorescence and Western blot experiments. Furthermore,

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12

Jerlström-Hultqvist, Jon, Elin Einarsson, and Staffan G. Svärd. "Stable Transfection of the Diplomonad Parasite Spironucleus salmonicida." Eukaryotic Cell 11, no. 11 (2012): 1353–61. http://dx.doi.org/10.1128/ec.00179-12.

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ABSTRACT Eukaryotic microbes are highly diverse, and many lineages remain poorly studied. One such lineage, the diplomonads, a group of binucleate heterotrophic flagellates, has been studied mainly due to the impact of Giardia intestinalis , an intestinal, diarrhea-causing parasite in humans and animals. Here we describe the development of a stable transfection system for use in Spironucleus salmonicida , a diplomonad that causes systemic spironucleosis in salmonid fish. We designed vectors in cassette format carrying epitope tags for localization (3×HA [where HA is hemagglutinin], 2× Escheric

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13

Nita, Akihiro, Akinobu Matsumoto, Ronghao Tang, et al. "A ubiquitin-like protein encoded by the “noncoding” RNA TINCR promotes keratinocyte proliferation and wound healing." PLOS Genetics 17, no. 8 (2021): e1009686. http://dx.doi.org/10.1371/journal.pgen.1009686.

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Although long noncoding RNAs (lncRNAs) are transcripts that do not encode proteins by definition, some lncRNAs actually contain small open reading frames that are translated. TINCR (terminal differentiation–induced ncRNA) has been recognized as a lncRNA that contributes to keratinocyte differentiation. However, we here show that TINCR encodes a ubiquitin-like protein that is well conserved among species and whose expression was confirmed by the generation of mice harboring a FLAG epitope tag sequence in the endogenous open reading frame as well as by targeted proteomics. Forced expression of t

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14

Jiang, Xiaofeng, Tracee S. Panetti, and Michael E. Bromberg. "The Cytoplasmic Domain of Tissue Factor Inhibits Migration and Enhances Adhesion of Human Breast Cancer Cells." Blood 106, no. 11 (2005): 1939. http://dx.doi.org/10.1182/blood.v106.11.1939.1939.

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Abstract Tissue factor (TF) is a 47 kDa transmembrane glycoprotein that when complexed with its cofactor, factor VIIa (FVIIa), initiates blood coagulation. Apart from hemostasis, TF has been shown to have roles in cellular signaling, development, inflammation, metastasis and angiogenesis. We showed previously that both the cytoplasmic and extracellular domains of TF are required for the full metastatic effect of TF. Recently, we showed that TF-FVIIa-FXa complex induces cellular signaling in human breast cancer cells and is associated with enhanced cell migration and prevention of apoptosis. Ho

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15

Walker, Michelle Portlance, and W. Ian Lipkin. "Characterization of the Nuclear Localization Signal of the Borna Disease Virus Polymerase." Journal of Virology 76, no. 16 (2002): 8460–67. http://dx.doi.org/10.1128/jvi.76.16.8460-8467.2002.

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ABSTRACT Borna disease virus (BDV) is a nonsegmented negative-strand RNA virus that replicates and transcribes its genome in the nucleus of infected cells. BDV proteins involved in replication and transcription must pass through the nuclear envelope to associate with the genomic viral RNA. The RNA-dependent RNA polymerase (L) of BDV is postulated to be the catalytic enzyme of replication and transcription. We demonstrated previously that BDV L localizes to the nucleus of BDV-infected cells and L-transfected cells. Nuclear localization of the protein presupposes the presence of a nuclear locali

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16

Maslennikova, Aleksandra, Dmitriy Komkov, Anastasia Zotova, and Dmitriy Mazurov. "Cell Surface-Expressed GPI-Anchored Peptides from the CHR Domain of gp41 Are Potent Inhibitors of HIV-1 Fusion." Proceedings 50, no. 1 (2020): 70. http://dx.doi.org/10.3390/proceedings2020050070.

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Current antiretroviral therapy efficiently suppresses viral replication but cannot eliminate latent HIV reservoirs. Moreover, the associated high costs, side effects, and drug resistance have stimulated a need for the development of alternative methods of HIV-1/AIDS treatment, such as peptide inhibitors or gene editing. Recently, we have developed Surface Oligopeptide knock-in for Rapid Target Selection (SORTS), a method for the rapid selection of CRISPR/Cas9 gene-edited cells via knock-in of the Flag and HA epitope tags embedded into the shortest GPI-protein, CD52. By targeting the capsid reg

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17

Cristea, Ileana M., Heather Rozjabek, Kelly R. Molloy, et al. "Host Factors Associated with the Sindbis Virus RNA-Dependent RNA Polymerase: Role for G3BP1 and G3BP2 in Virus Replication." Journal of Virology 84, no. 13 (2010): 6720–32. http://dx.doi.org/10.1128/jvi.01983-09.

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ABSTRACT Sindbis virus (SINV) is the prototype member of the Alphavirus genus, whose members cause severe human diseases for which there is no specific treatment. To ascertain host factors important in the replication of the SINV RNA genome, we generated a SINV expressing nsP4, the viral RNA-dependent RNA polymerase, with an in-frame 3×Flag epitope tag. Proteomic analysis of nsP4-containing complexes isolated from cells infected with the tagged virus revealed 29 associated host proteins. Of these, 10 proteins were associated only at a later time of infection (12 h), 14 were associated both ear

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18

Peters, Kathryn W., Juanjuan Qi, Simon C. Watkins, and Raymond A. Frizzell. "Syntaxin 1A inhibits regulated CFTR trafficking inXenopus oocytes." American Journal of Physiology-Cell Physiology 277, no. 1 (1999): C174—C180. http://dx.doi.org/10.1152/ajpcell.1999.277.1.c174.

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The cystic fibrosis transmembrane conductance regulator (CFTR) is an epithelial cell Cl channel, whose gating activity and membrane trafficking are controlled by cAMP/protein kinase A (PKA)-mediated phosphorylation. CFTR Cl currents are regulated also by syntaxin 1A (A. P. Naren, D. J. Nelson, W. W. Xie, B. Jovov, J. Pevsner, M. K. Bennett, D. J. Benos, M. W. Quick, and K. L. Kirk. Nature 390: 302–305, 1997), a protein best known for its role in membrane trafficking and neurosecretion. To examine the mechanism of syntaxin 1A inhibition, we expressed these proteins in Xenopusoocytes and monitor

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19

Bewarder, Moritz, Lorenz Thurner, Frank Neumann, et al. "BAR-Bodies: B-Cell Receptor Antigens As the Targeting Moiety of Antibodies in Substitution for the Variable Region of Heavy and Light Chains." Blood 132, Supplement 1 (2018): 2940. http://dx.doi.org/10.1182/blood-2018-99-115348.

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Abstract Background Chronic antigenic stimulation of the B-cell receptor (BCR) seems to play a critical role in the pathogenesis of B-cell lymphomas. We recently identified ARS2 and LRPAP1 as the autoantigenic targets of the B-cell receptors of approximately 25% of diffuse large B cell lymphomas (DLBCLs) of the ABC type and 45% of mantle cell lymphomas (MCLs), respectively. These BCR antigens can be used to target lymphoma cells in an approach we designated as BAR (B-cell receptor antigens for reverse targeting). The optimal therapeutic format BARs can be integrated in has yet to be found. Sin

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20

Bewarder, Moritz, Christina Körbel, Matthias W. Laschke, et al. "The B-Cell Receptor Antigen ARS2 Can be Integrated into a BAR-Body Format to Treat Diffuse Large B-Cell Lymphomas in Xenograft Mouse Models." Blood 134, Supplement_1 (2019): 2860. http://dx.doi.org/10.1182/blood-2019-131434.

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Chronic antigenic stimulation of the B-cell receptor (BCR) seems to play a critical role in the pathogenesis of B-cell lymphomas. We recently identified ARS2, LRPAP1 and Neurabin-I as the autoantigenic targets of the B-cell receptors of approximately 25% of diffuse large B cell lymphomas (DLBCLs) of the ABC type, 45% of mantle cell lymphomas (MCLs) and 66% of primary CNS lymphomas, respectively. These BCR antigens can be used to target lymphoma cells in an approach we designated as BAR (B-cell receptor antigens for reverse targeting). Since the most established approach to deliver therapeutic

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21

Stalder, Elizabeth S., Lauren H. Nagy, Pilar Batalla, Terrance M. Arthur, Nancy E. Thompson, and Richard R. Burgess. "The epitope for the polyol-responsive monoclonal antibody 8RB13 is in the flap-domain of the beta-subunit of bacterial RNA polymerase and can be used as an epitope tag for immunoaffinity chromatography." Protein Expression and Purification 77, no. 1 (2011): 26–33. http://dx.doi.org/10.1016/j.pep.2010.12.011.

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22

Dooher, Julia E., Ido Paz-Priel, Simone Houng, Albert S. Baldwin та Alan D. Friedman. "C/EBPα, C/EBPα Oncoproteins, or C/EBPβ Preferentially Bind NF-κB p50 Compared with p65 Via Conserved Residues In the C/EBP Basic Region". Blood 116, № 21 (2010): 708. http://dx.doi.org/10.1182/blood.v116.21.708.708.

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Abstract Abstract 708 Nuclear Factor-κB (NF-κB) is a key mediator of the inflammatory response and also inhibits apoptosis. Activation of NF-κB is prevalent in AML, Hodgkin's lymphoma and subsets of non-Hodgkin's lymphoma, as well as in multiple solid tumors. NF-κB p50 (NF-κB1) and p65 (Rel A) are the most prevalent NF-κB subunits. NF-κB p65 has 30–50 fold higher affinity than p50 for the cytoplasmic protein IκB, and so under basal conditions p50:p65 heterodimers are held in the cytoplasm whereas p50:p50 homodimers are found in the nucleus bound to chromatin where they repress NF-κB target gen

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23

Götzke, Hansjörg, Markus Kilisch, Markel Martínez-Carranza, et al. "The ALFA-tag is a highly versatile tool for nanobody-based bioscience applications." Nature Communications 10, no. 1 (2019). http://dx.doi.org/10.1038/s41467-019-12301-7.

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Abstract Specialized epitope tags are widely used for detecting, manipulating or purifying proteins, but often their versatility is limited. Here, we introduce the ALFA-tag, a rationally designed epitope tag that serves a remarkably broad spectrum of applications in life sciences while outperforming established tags like the HA-, FLAG®- or myc-tag. The ALFA-tag forms a small and stable α-helix that is functional irrespective of its position on the target protein in prokaryotic and eukaryotic hosts. We characterize a nanobody (NbALFA) binding ALFA-tagged proteins from native or fixed specimen w

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24

Imagawa, Toshifumi, Masahiko Ito, Mami Matsuda, et al. "Virus-like particles with FLAG-tagged envelope protein as a tetravalent dengue vaccine candidate." Scientific Reports 11, no. 1 (2021). http://dx.doi.org/10.1038/s41598-021-97038-4.

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AbstractThe global incidence of dengue, which is caused by dengue virus (DENV) infection, has grown dramatically in recent decades and secondary infection with heterologous serotype of the virus may cause severe symptoms. Efficacious dengue vaccines should be able to provide long-lasting immunity against all four DENV serotypes simultaneously. In this study, we constructed a novel vaccine platform based on tetravalent dengue virus-like particles (DENV-LPs) in which envelope (E) protein carried a FLAG tag sequence at the position located not only in the exterior loop on the protruding domain bu

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25

Yang, Hui, Yu Wang, and Chengjin Ye. "Rapid Generation of Attenuated Infectious Bursal Disease Virus from Dual-Promoter Plasmids by Reduction of Viral Ribonucleoprotein Activity." Journal of Virology 94, no. 7 (2020). http://dx.doi.org/10.1128/jvi.01569-19.

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ABSTRACT Infectious bursal disease virus (IBDV) of the Birnaviridae family leads to immunosuppression of young chickens by destroying B cells in the bursa of Fabricius (BFs). Given the increasing number of variant IBDV strains, we urgently require a method to produce attenuated virus for vaccine development. To accomplish this goal, the dual-promoter plasmids in which the RNA polymerase II and RNA polymerase I (Pol I) promoters were placed upstream of the IBDV genomic sequence, which was followed by mouse Pol I terminator and a synthetic polyadenylation signal, were developed for rapid generat

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26

Carakushansky, M., AJ Whatmore, PE Clayton, et al. "A new missense mutation in the growth hormone-releasing hormone receptor gene in familial isolated GH deficiency." European Journal of Endocrinology, January 1, 2003, 25–30. http://dx.doi.org/10.1530/eje.0.1480025.

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OBJECTIVE: Mutations in the GH-releasing hormone (GHRH) receptor (GHRHR) gene (GHRHR) cause autosomal recessive familial isolated GH deficiency (IGHD). We searched for GHRHR mutations in two siblings with IGHD type IB and a history of parental consanguinity. DESIGN: We analyzed peripheral genomic DNA of an index patient. After identifying a novel mutation in the GHRHR, we performed functional studies in order to confirm that the mutation causes receptor malfunction. METHODS: The entire GHRHR was analyzed in the index case by denaturing gradient gel electrophoresis. Abnormally migrating bands w

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