摘要
根据已发表的弓形虫主要表面抗原（ Ｐ３０）的基因序列，设计合成了一对引物，通过聚合酶链反应，扩增出 Ｐ３０ 基因，将 Ｐ３０ 基因克隆入表达质粒ｐ Ｇ Ｅ Ｍ Ｅ Ｘ１ 中，经酶切鉴定后，阳性重组质粒转化宿主菌 Ｊ Ｍ １０９（ Ｄ Ｅ３ ），宿主菌经 Ｉ Ｐ Ｔ Ｇ 诱导表达后，产物进行 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ和 Ｗ ｅｓｔｅｒｎｂｌｏｔ分析。结果显示， Ｐ３０ 基因以融合蛋白的形式表达，表达产物具有特异的免疫反应性。
According to the published gene sequence of the major surface antigen (p30) of Toxoplasma gondii , a pair of primers was designed and synthesized. The coding sequences of p30 gene were amplified by polymerase chain reaction and cloned into the expression plasmid pGEMEX 1. The recombinant plasmids were analysed by restriction enzyme. The positive recombinant plasmids were transformed into the host bacteria JM109(DE 3). The host bacteria was induced by IPTG. SDS polyacrylamide gel electrophoresis and immunoblot analysis of total cellular protein from JM109(DE 3) were performed to detect the p30 gene product. The results shown that the p30 gene cloned into the expression plasmid pGEMEX 1 could express in JM109(DE 3) as a fusion protein. The expression product had immunogenicity.
出处
《中国寄生虫病防治杂志》
CSCD
1999年第3期192-194,共3页
Chinese Journal of Parasitic Disease Control
基金
山东省科委和计生委资助
关键词
弓形虫
表面抗原
基因克隆
基因表达
Toxoplasma gondii , major surface antigen, polymerase chain reaction,gene cloning,gene expression
