痢疾菌体内诱导基因表达文库的构建  

Construction of a fusion gene expression library screening in vivo-induced genes in Shigella flexneri 2a

作　　者：史兆兴[1] 王恒樑[1] 冯尔玲[1] 姚潇[2] 廖翔[1] 黄留玉[1] 苏国富[1] 黄翠芬[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071 [2]西安交通大学环境与化学工程学院,陕西西安710049

出　　处：《细胞与分子免疫学杂志》2001年第5期432-433,共2页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的构建筛选痢疾菌福氏2a体内诱导基因的融合基因表达文库。方法以自杀载体pGP704为骨架,用氯霉素乙酰转移酶基因作为报告基因,构建了用于筛选体内诱导基因的载体pGP-cat。把痢疾菌福氏2a基因组DNA的随机酶切片段0.6-1kb亚克隆到pGPcat载体报告基因上游的BglⅡ位点,构建了融合基因文库。结果与痢疾菌福氏2a2457T结合转移后,获得融合基因表达文库。以氯霉素为选择标记,经过体外初步筛选,得到3.2%的具有氯霉素抗性的菌株。结论构建的融合基因表达文库适用于筛选福氏2a痢疾菌的体内诱导基因。Aim To construct a fusion gene expressio n library screening in vivo-induced genes in Flex shigella.Methods Based on the suicide vector pGP704,a new screening in vivo-indu ced genes vector pGPcat was constructed with chloramphenicol a cetyltransferase(cat)gene as a report gene.The random Shigella flexneri 2a DNA fragments(0.6-1kb),obtained by partial Sau3A1restriction digestion,were subcloned into t he unique Bgl Ⅱsite of pGPcat,5'to the promoterless reporter gene.And the fusion gene library was constructed.Results The fusion gene expression library was obtained by mating into Shigella flexneri strain 2457T.3.2%Cm r strains were obtained by screening i n vitro with chloramphenicol as the selectable marker.Conclusion The expression library is suitable f or screening in vivo-induced genes of Shigella flexneri 2a.

关 键 词：氯霉素乙酰转移酶 体内诱导基因 融合基因表达文库 福氏2a痢疾菌 

分 类 号：R378.25[医药卫生—病原生物学]