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弓形虫主要表面抗原基因( P30)大肠杆菌中以融合蛋白形式的初步表达(英文)
PRIMARY EXPRESSION OF THE MAJOR SURFACE ANTIGEN (P30) GENE OF TOXOPLASMA GONDII AS A FUSION PROTEIN IN E. COLI *
【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。
【Abstract】 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.
【Key words】 Toxoplasma gondii; major surface antigen; polymerase chain reaction; gene cloning; gene expression;
- 【文献出处】 中国寄生虫病防治杂志 ,CHINESE JOURNAL OF PARASITIC DISEASE CONTROL , 编辑部邮箱 ,1999年03期
- 【分类号】R382.5
- 【被引频次】3
- 【下载频次】22