Murata T, Xuan X, Otsuka H
Department of Global Agricultural Sciences, Graduate School of Agricultural and Life Science, The University of Tokyo, Japan.
J Vet Med Sci. 1999 May;61(5):453-7. doi: 10.1292/jvms.61.453.
Bovine herpesvirus-1 (BHV-1) has been used as a vector of live recombinant vaccines for cattle which express the genes of other pathogens. Because of the importance of the choice of the promoter which allows the efficient expression of the foreign genes in the BHV-1 vector, we compared the relative efficacy of various promoters integrated in the BHV-1 genome. The promoter sequences of the BHV-1 thymidine kinase (tk), gB, gC, SV40 early, and pseudorabies virus (PRV) immediate early (IE) genes were placed at the upstream of the open reading frame of the chloramphenycol acetyl transferase (CAT) gene and the promoter-CAT sequences were integrated into the tk gene of BHV-1 by homologous recombination. The promoter activity was assayed by measuring the CAT activity in the extracts of Madin Darby bovine kidney (MDBK) cells infected with the recombinant BHV-1. The PRV IE promoter was activated earlier and maintained at a higher level activity than the BHV-1 gB or gC promoters throughout the most of the growth phase of BHV-1. At the late phase, however, the activities of the BHV-1 gB and gC promoters reached the higher level. The BHV-1 tk promoter activity was low and the SV40 early promoter was hardly activated when integrated into the BHV-1 genome. promoter, recombinant BHV-1.
牛疱疹病毒1型（BHV-1）已被用作表达其他病原体基因的牛用重组活疫苗载体。由于启动子的选择对于在BHV-1载体中高效表达外源基因至关重要，我们比较了整合在BHV-1基因组中的各种启动子的相对效力。将BHV-1胸苷激酶（tk）、gB、gC、SV40早期和伪狂犬病病毒（PRV）立即早期（IE）基因的启动子序列置于氯霉素乙酰转移酶（CAT）基因开放阅读框的上游，并通过同源重组将启动子-CAT序列整合到BHV-1的tk基因中。通过测量感染重组BHV-1的马迪达比牛肾（MDBK）细胞提取物中的CAT活性来测定启动子活性。在BHV-1的大部分生长阶段，PRV IE启动子比BHV-1 gB或gC启动子更早被激活并维持在更高水平的活性。然而，在后期，BHV-1 gB和gC启动子的活性达到更高水平。当整合到BHV-1基因组中时，BHV-1 tk启动子活性较低，SV40早期启动子几乎不被激活。启动子，重组BHV-1。
