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.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2001年第5期432-433,共2页
Chinese Journal of Cellular and Molecular Immunology
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
chloramphenicol acetyltransfera se
in vivo-induced genes
fusion gene expression library
shigella flexneri 2a
