Vaccinia virus expression vector: coexpression of beta-galactosidase provides visual screening of recombinant virus plaques.
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TLDR
Each protocol was tested by constructing vaccinia virus recombinants that express hepatitis B virus surface antigen by constructing plasmid coexpression vector that directs the insertion of a foreign gene of interest together with the Escherichia coli beta-galactosidase (beta gal) gene into the thymidine kinase (TK) locus of the vaccinations genome.Abstract:
We constructed a plasmid coexpression vector that directs the insertion of a foreign gene of interest together with the Escherichia coli beta-galactosidase (beta gal) gene into the thymidine kinase (TK) locus of the vaccinia virus genome. Tissue culture cells that had been infected with vaccinia virus were transfected with a plasmid vector containing a foreign gene. TK- recombinants could be selected by a plaque assay on TK- cells in the presence of 5-bromodeoxyuridine and distinguished from spontaneous TK- mutants by the addition of a beta-gal indicator to the agarose overlay. Plaques that expressed beta-gal stained dark blue within several hours at 37 degrees C. Alternatively, TK- selection could be eliminated, and recombinant plaques could be readily identified solely by their blue color. The reverse procedure, in which the starting virus expresses beta-gal (i.e., forms blue plaques) and the desired recombinant has deleted the entire beta-gal gene (i.e., forms white plaques), is another alternative. Each protocol was tested by constructing vaccinia virus recombinants that express hepatitis B virus surface antigen.read more
Citations
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Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase
TL;DR: The vaccinia/T7 hybrid virus forms the basis of a simple, rapid, widely applicable, and efficient mammalian expression system.
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Virus-specific CD8+ cytotoxic T-lymphocyte activity associated with control of viremia in primary human immunodeficiency virus type 1 infection.
TL;DR: HIV-1-specific CTL activity is a major component of the host immune response associated with the control of virus replication following primary HIV-1 infection and have important implications for the design of antiviral vaccines.
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A novel influenza A virus mitochondrial protein that induces cell death
Weisan Chen,Paul A. Calvo,Daniela Malide,James P. Gibbs,Ulrich Schubert,Igor Bacik,Sameh Basta,Robert E. O'Neill,Jeanne Schickli,Peter Palese,Peter Henklein,Jack R. Bennink,Jonathan W. Yewdell +12 more
TL;DR: It is proposed that PB1-F2 functions to kill host immune cells responding to influenza virus infection, and influenza viruses with targeted mutations that interfere with PB1/F2 expression induce less extensive apoptosis in human monocytic cells than those with intact PB1 -F2.
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CD1 recognition by mouse NK1+ T lymphocytes
Albert Bendelac,Albert Bendelac,Olivier Lantz,Mary E. Quimby,Jonathan W. Yewdell,Jack R. Bennink,Randy R. Brutkiewicz +6 more
TL;DR: In mice, an entire subset of alpha beta thymocytes with a unique phenotype was found to be CD1-specific, providing evidence that CD1 has a general role in selecting and interacting with specialized alpha beta T cells.
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A phagosome-to-cytosol pathway for exogenous antigens presented on MHC class I molecules
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References
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Book
Experiments in molecular genetics
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Journal ArticleDOI
A system for shotgun DNA sequencing.
TL;DR: A multipurpose cloning site has been introduced into the gene for beta-galactosidase on the single-stranded DNA phage M13mp2 with the use of synthetic DNA and two restriction endonuclease cleavage sites in the viral gene II were removed by single base-pair mutations.
Journal ArticleDOI
Vaccinia virus: a selectable eukaryotic cloning and expression vector
TL;DR: These studies demonstrate the use ofvaccinia virus as a selectable cloning and expression vector, confirm the map location of the vaccinia virus TK gene, and provide initial information regarding the location of vacciniairus transcriptional regulatory sequences.
Journal ArticleDOI
General method for production and selection of infectious vaccinia virus recombinants expressing foreign genes.
TL;DR: The production and selection of infectious vaccinia virus recombinants expressing foreign genes was facilitated by the construction of plasmid vectors and Infectious recombinant viruses expressing the procaryotic enzyme chloramphenicol acetyltransferase were constructed to optimize the system.
Book ChapterDOI
Beta-galactosidase gene fusions for analyzing gene expression in Escherichia coli and yeast
TL;DR: In vitro methods and some recently developed β -galactosidase gene fusion vectors are described, which can be used not only to measure gene expression and regulation, but also to isolate mutations and additional gene fusions.
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