Construction of canine herpesvirus vector expressing foreign genes using a lacZ-TK gene cassette as a double selectional marker

An improved method for constructing canine herpesvirus (CHV) recombinants expressing foreign genes by using the lacZ-TK gene cassette as a double selectional marker was developed. A recombinant CHV carrying the lacZ-TK gene at a targeted gene locus was constructed and used as a parental virus for ge...

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Bibliographic Details
Published in:Virus genes 1998-01, Vol.17 (1), p.25-32
Main Authors: Xuan, X, Nishikawa, Y, Takashima, Y, Tuchiya, K, Ueda, S, Yokoyama, N, Maeda, K, Mikami, T, Otsuka, H
Format: Article
Language:English
Subjects:
Animals
Biomarkers
Cell Line
Cloning, Molecular
DNA, Viral - analysis
DNA, Viral - genetics
Dogs
Gene Expression Regulation, Viral
Genetic Vectors - genetics
Herpesvirus 1, Canid - genetics
Lac Operon - genetics
Plaques
Rabies
Recombinant Fusion Proteins - analysis
Recombinant Fusion Proteins - genetics
Recombinants
Thymidine Kinase - genetics
TK gene
Transgenes - genetics
Viruses
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Summary:An improved method for constructing canine herpesvirus (CHV) recombinants expressing foreign genes by using the lacZ-TK gene cassette as a double selectional marker was developed. A recombinant CHV carrying the lacZ-TK gene at a targeted gene locus was constructed and used as a parental virus for generating new recombinants. The parental virus formed blue plaques and was sensitive to TK-specific drugs, while newly generated recombinants, in which the lacZ-TK gene was replaced with the desired foreign gene, become both resistant to the TK-specific drugs and formed white plaques. Recombinants were isolated by using the combination of drug selection and color selection. This improved method allows construction of recombinant CHV with great ease, because the drug selection can enrich the frequency of recombinant CHV from 0.01-0.1% to 10-80%. This method was employed to construct a recombinant CHV that expressed rabies virus (RV) glycoprotein (G protein).
ISSN:0920-8569
1572-994X
DOI:10.1023/a:1008096832738