中国动物传染病学报Issue(4)：17-22,6.
乙型脑炎病毒基因组cDNA克隆在宿主菌中不稳定因子的初步研究
PRELIMINARY STUDY ON GENETIC INSTABILITY OF JAPANESES ENCEPHALITIS VIRUS CDNACLONE TRANSFORMED IN ESCHERICHIA COLI
摘要
Abstract
In the present study, a series of fragments from 1 to 2913 bp were generated from Japanese encephalitis virus (JEV) HEN0701genome in order to explore factors affecting genetic instability of cDNA clone when propagated in E. coli. These cDNA fragments were obtained using either PCR starting at 5' terminus or restriction enzyme digestion at different positiosn of 3' terminus. Each fragment was cloned into pCR-BluntII-TOPO vector and transformed into E. coli cultures, then were incubated at 37℃. Fragments 73 to 2913 bp, 97 to 2913 bp, 355 to 2913 bp and 1 to 933 bp, 1 to 1275 bp showed stability during passages of the transformed bacteria. However, recombinant bacteria containing fragments of 39 to 2913 bp, 54 to 2913 bp and 1 to 1800 bp, 1 to 1971 bp did not grow at 37℃. In addition, fragment 1 to 1600 bp of JEV genome was passaged in transformed bacteria but mutation occurred. The above results indicated that the fragment 73 to 118 bp affected the stability of JEV cDNA clone. The upstream sequences at 54 to 73 bp might be involve in this process as well. The results also suggest that toxic protein coding sequence might exist between 1275 to 1600 bp of JEV genome.关键词
乙型脑炎病毒/遗传不稳定性/潜在启动子/毒性蛋白编码序列位置Key words
Japanese encephalitis virus (JEV)/genetic instability/potential prokaryotic promoter/toxic protein coding sequence position分类
农业科技引用本文复制引用
郑旭晨,郑浩,郭亦非,刘飞,梁超,童武,单同领,于海,童光志..乙型脑炎病毒基因组cDNA克隆在宿主菌中不稳定因子的初步研究[J].中国动物传染病学报,2014,(4):17-22,6.基金项目
上海市自然基金项目（11ZR1446200）；国家自然基金青年基金项目（31201917）；上海市重点科技攻关项目（09391910800）；中央级公益性科研院所基本科研业务费项目 （）
