Construction of Canine Herpesvirus Vector Expressing Foreign Genes Using a lacZ-TK Gene Cassette as a Double Selectional Marker |
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| Authors: | Xuan Xuenan Nishikawa Yoshifumi Takashima Yasuhiro Tuchiya Kotaro Ueda Susumu Yokoyama Naoaki Maeda Ken Mikami Takeshi Otsuka Haruki |
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| Affiliation: | (1) Department of Global Agricultural Science, Graduate School of Agricultural Science, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan;(2) Nippon Institute for Biological Science, 2221-1 Shin-machi, Ome, Tokyo 198, Japan;(3) Department of Veterinary Microbiology, Graduate School of Agricultural Science, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan;(4) Department of Veterinary Microbiology, Faculty of Agriculture, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753, Japan;(5) Present address: The Research Center for Protozoan Molecular Immunology, Obihiro University of Agriculture and Veterinary Medicine, 2-13 Nishi, Inada-cho, Obihiro, Hokkaido 080, Japan |
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| Abstract: | 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). This revised version was published online in July 2006 with corrections to the Cover Date. |
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| Keywords: | canine herpesvirus lacZ-TK recombinant rabies virus glycoprotein |
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