Chowdhury S I
Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan 66506, USA.
Vet Microbiol. 1996 Sep;52(1-2):13-23. doi: 10.1016/0378-1135(96)00043-0.
A recombinant bovine herpesvirus type 1 (BHV-1) virus (Gal-TK) has been constructed. The Gal-TK virus contains a chimeric reporter/marker gene coding for bacterial beta-galactosidase (beta-gal gene) that was inserted stably within the viral TK gene. This resulted in inactivation of the TK gene. The beta-gal gene is under the regulation of a strong, human cytomegalovirus-immediate early (HCMV-IE) promoter and is expressed as an authentic viral-coded gene. Even though the one-step growth kinetics of the recombinant and parent viruses were similar, the recombinant virus yielded less than the parent virus on Madin-Darby bovine kidney cells. After intranasal inoculation, the engineered virus was virtually avirulent for colostrum-deprived new-born calves. Similar to the parent virus, the recombinant virus replicated in the upper respiratory tract of calves, but the amount of progeny viruses produced was reduced significantly. The progeny viruses recovered from nasal swabs of animals inoculated with the recombinant and the Cooper strains of BHV-1 were easily distinguishable based on the beta-gal marker.
已构建出一种重组牛疱疹病毒1型（BHV-1）病毒（Gal-TK）。Gal-TK病毒包含一个嵌合报告基因/标记基因，其编码细菌β-半乳糖苷酶（β-gal基因），该基因稳定插入病毒胸苷激酶（TK）基因内。这导致TK基因失活。β-gal基因受强人类巨细胞病毒立即早期（HCMV-IE）启动子调控，并作为一个真实的病毒编码基因表达。尽管重组病毒和亲本病毒的一步生长动力学相似，但重组病毒在马-达二氏牛肾细胞上的产量低于亲本病毒。经鼻内接种后，工程病毒对初乳缺乏的新生犊牛几乎无毒力。与亲本病毒相似，重组病毒在犊牛的上呼吸道中复制，但产生的子代病毒数量显著减少。根据β-gal标记，从接种重组BHV-1和库珀株的动物鼻拭子中回收的子代病毒很容易区分。
