Structure and stability of mRNA synthesized by vaccinia virus-encoded bacteriophage T7 RNA polymerase in mammalian cells

1989 ◽  
Vol 206 (2) ◽  
pp. 333-348 ◽  
Author(s):  
Thomas R. Fuerst ◽  
Bernard Moss
Keyword(s):  
Rna Polymerase ◽  
Vaccinia Virus ◽  
Mammalian Cells ◽  
T7 Rna Polymerase ◽  
Bacteriophage T7 ◽  
Structure And Stability

scholarly journals Expression of bacteriophage T7 RNA polymerase in avian and mammalian cells by a recombinant fowlpox virus

1996 ◽  
Vol 77 (5) ◽  
pp. 963-967 ◽  
Author(s):  
P. Britton ◽  
P. Green ◽  
S. Kottier ◽  
K. L. Mawditt ◽  
Z. Penzes ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Mammalian Cells ◽  
T7 Rna Polymerase ◽  
Bacteriophage T7 ◽  
Fowlpox Virus ◽  
Recombinant Fowlpox Virus

scholarly journals Use of a hybrid vaccinia virus-T7 RNA polymerase system for expression of target genes.

1987 ◽  
Vol 7 (7) ◽  
pp. 2538-2544 ◽  
Author(s):  
T R Fuerst ◽  
P L Earl ◽  
B Moss
Keyword(s):  
Rna Polymerase ◽  
Vaccinia Virus ◽  
Target Gene ◽  
Target Genes ◽  
Expression System ◽  
Recombinant Vaccinia Virus ◽  
T7 Rna Polymerase ◽  
Bacteriophage T7 ◽  
Virus Surface ◽  
Recombinant Vaccinia

A novel expression system based on coinfection of cells with two recombinant vaccinia viruses has been developed. One recombinant vaccinia virus contained the bacteriophage T7 RNA polymerase gene under control of a vaccinia virus promoter. The second recombinant vaccinia virus contained a target gene of choice flanked by bacteriophage T7 promoter and termination sequences. Maximum expression of the target gene occurred when cells were infected with 10 PFU of each recombinant virus. Although T7 RNA polymerase synthesis began shortly after infection, the target gene was not expressed until late times and was largely inhibited when DNA replication was blocked. Target gene transcripts were analyzed by agarose gel electrophoresis and had the predicted size. With this system, Escherichia coli beta-galactosidase, hepatitis B virus surface antigen, and human immunodeficiency virus envelope proteins were made. In each case, the level of synthesis was greater than had previously been obtained with the more conventional recombinant vaccinia virus expression system.

Use of a hybrid vaccinia virus-T7 RNA polymerase system for expression of target genes

1987 ◽  
Vol 7 (7) ◽  
pp. 2538-2544
Author(s):  
T R Fuerst ◽  
P L Earl ◽  
B Moss
Keyword(s):  
Rna Polymerase ◽  
Vaccinia Virus ◽  
Target Gene ◽  
Target Genes ◽  
Expression System ◽  
Recombinant Vaccinia Virus ◽  
T7 Rna Polymerase ◽  
Bacteriophage T7 ◽  
Virus Surface ◽  
Recombinant Vaccinia

A novel expression system based on coinfection of cells with two recombinant vaccinia viruses has been developed. One recombinant vaccinia virus contained the bacteriophage T7 RNA polymerase gene under control of a vaccinia virus promoter. The second recombinant vaccinia virus contained a target gene of choice flanked by bacteriophage T7 promoter and termination sequences. Maximum expression of the target gene occurred when cells were infected with 10 PFU of each recombinant virus. Although T7 RNA polymerase synthesis began shortly after infection, the target gene was not expressed until late times and was largely inhibited when DNA replication was blocked. Target gene transcripts were analyzed by agarose gel electrophoresis and had the predicted size. With this system, Escherichia coli beta-galactosidase, hepatitis B virus surface antigen, and human immunodeficiency virus envelope proteins were made. In each case, the level of synthesis was greater than had previously been obtained with the more conventional recombinant vaccinia virus expression system.

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