Tn7-mediated introduction of DNA into bacmid-cloned pseudorabies virus genome for rapid construction of recombinant viruses

2007 ◽  
Vol 22 (4) ◽  
pp. 316-325 ◽  
Author(s):  
Fang-fang Zhuan ◽  
Zhen-feng Zhang ◽  
Di-ping Xu ◽  
Yan-hong Si ◽  
Han-Zhong Wang ◽  
...  
Keyword(s):  
Pseudorabies Virus ◽  
Virus Genome ◽  
Recombinant Viruses ◽  
Rapid Construction

scholarly journals Complete, Annotated Sequence of the Pseudorabies Virus Genome

2004 ◽  
Vol 78 (4) ◽  
pp. 2166-2166 ◽  
Author(s):  
Barbara G. Klupp ◽  
Christoph J. Hengartner ◽  
Thomas C. Mettenleiter ◽  
Lynn W. Enquist
Keyword(s):  
Pseudorabies Virus ◽  
Virus Genome

A simple and rapid approach to manipulate pseudorabies virus genome by CRISPR/Cas9 system

2015 ◽  
Vol 37 (6) ◽  
pp. 1265-1272 ◽  
Author(s):  
Aotian Xu ◽  
Chao Qin ◽  
Yue Lang ◽  
Mingyue Wang ◽  
Mengyang Lin ◽  
...  
Keyword(s):  
Pseudorabies Virus ◽  
Virus Genome

scholarly journals Viral Bacterial Artificial Chromosomes: Generation, Mutagenesis, and Removal of Mini-F Sequences

2012 ◽  
Vol 2012 ◽  
pp. 1-14 ◽  
Author(s):  
B. Karsten Tischer ◽  
Benedikt B. Kaufer
Keyword(s):  
Dna Sequences ◽  
Rna Virus ◽  
Virus Genome ◽  
Virus Species ◽  
Bacterial Artificial Chromosomes ◽  
Nucleotide Substitutions ◽  
Recombinant Viruses ◽  
Artificial Chromosomes ◽  
Dna And Rna ◽  
Virus Genomes

Maintenance and manipulation of large DNA and RNA virus genomes had presented an obstacle for virological research. BAC vectors provided a solution to both problems as they can harbor large DNA sequences and can efficiently be modified using well-established mutagenesis techniques inEscherichia coli. Numerous DNA virus genomes of herpesvirus and pox virus were cloned into mini-F vectors. In addition, several reverse genetic systems for RNA viruses such as members ofCoronaviridaeandFlaviviridaecould be established based on BAC constructs. Transfection into susceptible eukaryotic cells of virus DNA cloned as a BAC allows reconstitution of recombinant viruses. In this paper, we provide an overview on the strategies that can be used for the generation of virus BAC vectors and also on systems that are currently available for various virus species. Furthermore, we address common mutagenesis techniques that allow modification of BACs from single-nucleotide substitutions to deletion of viral genes or insertion of foreign sequences. Finally, we review the reconstitution of viruses from BAC vectors and the removal of the bacterial sequences from the virus genome during this process.

scholarly journals A novel approach to propagate flavivirus infectious cDNA clones in bacteria by introducing tandem repeat sequences upstream of virus genome

2014 ◽  
Vol 95 (7) ◽  
pp. 1493-1503 ◽  
Author(s):  
Szu-Yuan Pu ◽  
Ren-Huang Wu ◽  
Ming-Han Tsai ◽  
Chi-Chen Yang ◽  
Chung-Ming Chang ◽  
...  
Keyword(s):  
Growth Kinetics ◽  
Tandem Repeat ◽  
Viral Genome ◽  
Vaccine Development ◽  
Virus Genome ◽  
Cdna Clones ◽  
Recombinant Viruses ◽  
Repeat Sequences ◽  
Tandem Repeat Sequences ◽  
Kinetics Of

Despite tremendous efforts to improve the methodology for constructing flavivirus infectious cDNAs, the manipulation of flavivirus cDNAs remains a difficult task in bacteria. Here, we successfully propagated DNA-launched type 2 dengue virus (DENV2) and Japanese encephalitis virus (JEV) infectious cDNAs by introducing seven repeats of the tetracycline-response element (7×TRE) and a minimal cytomegalovirus (CMVmin) promoter upstream of the viral genome. Insertion of the 7×TRE-CMVmin sequence upstream of the DENV2 or JEV genome decreased the cryptic E. coli promoter (ECP) activity of the viral genome in bacteria, as measured using fusion constructs containing DENV2 or JEV segments and the reporter gene Renilla luciferase in an empty vector. The growth kinetics of recombinant viruses derived from DNA-launched DENV2 and JEV infectious cDNAs were similar to those of parental viruses. Similarly, RNA-launched DENV2 infectious cDNAs were generated by inserting 7×TRE-CMVmin, five repeats of the GAL4 upstream activating sequence, or five repeats of BamHI linkers upstream of the DENV2 genome. All three tandem repeat sequences decreased the ECP activity of the DENV2 genome in bacteria. Notably, 7×TRE-CMVmin stabilized RNA-launched JEV infectious cDNAs and reduced the ECP activity of the JEV genome in bacteria. The growth kinetics of recombinant viruses derived from RNA-launched DENV2 and JEV infectious cDNAs displayed patterns similar to those of the parental viruses. These results support a novel methodology for constructing flavivirus infectious cDNAs, which will facilitate research in virology, viral pathogenesis and vaccine development of flaviviruses and other RNA viruses.

Equalization of the inverted repeat sequences of the pseudorabies virus genome by intermolecular recombination

Virology ◽  
1984 ◽  
Vol 132 (2) ◽  
pp. 303-314 ◽  
Author(s):  
Tamar Ben-Porat ◽  
Anne Deatly ◽  
Ruth Ann Veach ◽  
Mayme L. Blankenship
Keyword(s):  
Inverted Repeat ◽  
Pseudorabies Virus ◽  
Virus Genome ◽  
Repeat Sequences

Transcription from the pseudorabies virus genome during latent infection

1988 ◽  
Vol 98 (1-2) ◽  
pp. 99-106 ◽  
Author(s):  
D. L. Rock ◽  
W. A. Hagemoser ◽  
F. A. Osorio ◽  
H. A. McAllister
Keyword(s):  
Pseudorabies Virus ◽  
Latent Infection ◽  
Virus Genome

scholarly journals Comparison of in Vivo Reactivation, in Vitro Reactivation, and Polymerase Chain Reaction for Detection of Latent Pseudorabies Virus Infection in Swine

1993 ◽  
Vol 5 (4) ◽  
pp. 505-509 ◽  
Author(s):  
Susan L. Brockmeier ◽  
Kelly M. Lager ◽  
William L. Mengeling
Keyword(s):  
Polymerase Chain Reaction ◽  
Pseudorabies Virus ◽  
Virus Genome ◽  
Explant Culture ◽  
Trigeminal Ganglia ◽  
Chain Reaction ◽  
Successful Method ◽  
Polymerase Chain

The following methods were compared for their ability to detect latent pseudorabies virus in 24 pigs that had been experimentally infected with virulent pseudorabies virus: 1) in vivo reactivation by dexamethasone administration, 2) in vitro reactivation by 5 different techniques of explant culture or cocultivation of trigeminal ganglia, and 3) detection of pseudorabies virus genome in tissue digests of tonsils or trigeminal ganglia using the polymerase chain reaction. Reactivation of pseudorabies virus by administration of dexamethasone was attempted in 12 of 24 pigs in an effort to determine if this procedure would affect the detection of latent pseudorabies virus by any of the subsequent in vitro methods. Detection of latent virus by the polymerase chain reaction with trigeminal ganglia was the most successful method (24/24 were positive). The next most successful method was in vivo reactivation through the administration of dexamethasone (10/12 [83%] were positive). Only 1 in vitro reactivation technique, cocultivation involving digestion of the trigeminal ganglia with trypsin and collagenase and the addition of a hypomethylating agent to the medium, yielded positive results (5/24 [21%] were positive). The polymerase chain reaction performed on tissue digests of tonsils was much less effective (2/24 [8%] were positive) than it was with trigeminal ganglia. Reactivation by dexamethasone did not appear to have any effect on the subsequent detection of latency by any of the methods tested.

Sign in / Sign up

Export Citation Format

Share Document