中国人兽共患病杂志2005，Vol.21Issue(12)：1089-1093,5.
刚地弓形虫P30(SAG1)重组抗原的高效表达和体外纯化
Over-expression and purification of the recombinant p30 antigen of Toxoplasma gondii
张佃波 1宰德富 1公茂庆 1李瑾 1魏庆宽 1崔勇 1黄炳成 1刘克义1
作者信息
- 1. 山东省寄生虫病防治研究所,济宁,272033
- 折叠
摘要
Abstract
To provide the basis for preparation of diagnostic kits and vaccines in Toxoplasma gondii infection, the gene coding for the qualified recombinant p30 protein (SAG1) of this parasite was amplified by PCR, and the amplified gene was cloned into prokaryotic expression vector pET-30a(+) to construct the recombinant plasmid, and then transformed to E.coli DH5α. The positive recombinant plasmid was screened by PCR and double enzymes digestion, and the nucleotide sequence of p30 gene was determined by automated DNA sequencing. Meanwhile, the identified recombinant plasmid was transformed to E.coli BL21(DE3) with the expression of p30 on bacteria induced by IPTG and the expressed protein was identified by SDS-PAGE. The protein obtained was then further purified and refolded, and its biological activity was checked by Western blotting. It was shown that the size of the amplified gene was 750 bp with molecular weight of 30 ku, and this protein could specifically react with monoclonal antibody against p30 protein.关键词
刚地弓形虫/P30基因/融合蛋白纯化Key words
T. gondii/P30 gene/fusion protein purification分类
医药卫生引用本文复制引用
张佃波,宰德富,公茂庆,李瑾,魏庆宽,崔勇,黄炳成,刘克义..刚地弓形虫P30(SAG1)重组抗原的高效表达和体外纯化[J].中国人兽共患病杂志,2005,21(12):1089-1093,5.
