弓形虫ROP2基因的克隆及原核表达
loning and Protoaryotic expression of the Toxoplasma gondii ROP2 Gene
应用PCR技术从刚地弓形虫( Toxoplasma gondii )RH株的基因组DNA中扩增编码ROP2(rhpotry protein 2)的部分基因，构建pGEX-KG-ROP2重组表达质粒，经酶切、PCR及DNA测序鉴定后，将阳性质粒转入E.coliBL21CodonPlus中在IPTG诱导下表达，表达产物用SDS-PAGE和Western-blot分析鉴定。结果表明，扩增的ROP2基因与GenBank上发表的相应基因序列的同源性达99.9 %，该基因可以在大肠杆菌中高效表达，表达的ROP2融合蛋白表观分子量约为64 KD，可被兔抗弓形虫免疫血清识别。本研究为下一步利用重组蛋白研制弓形虫病的诊断试剂盒和基因工程疫苗奠定基础。
he truncated ROP2 gene was obtained by amplification from genomic DNA of Toxoplasma gondii RH strain and cloned into the plasmid pGEX-KG. The constructed recombinant plasmid was identified by enzyme digesting, PCR and sequencing .Then the target recombinant plasmid was transferred into E.coli BL21Codonplus susceptible cell, the expression of the recombinant pGEX-KG-ROP2 was induced by IPTG. The expression protein were analysed by SDS-PAGE and by Western-blot. The results showed that the sequence identities are 99.9 % between amplified ROP2 gene and that from Genebank. The molecular weight of fused ROP2 was about 64 KD by SDS-PAGE. It could be specifically recognized by immune serum against Toxoplasma gondii from. Rabbit.
聂浩、姚宝安、周艳琴、刘琴、江涛、赵俊龙
基础医学分子生物学生物工程学
刚地弓形虫棒状体蛋白ROP2基因克隆原核表达
oxoplasma gondiiRhoptry protein ROP2Gene cloneProtoaryotic expression
聂浩,姚宝安,周艳琴,刘琴,江涛,赵俊龙.弓形虫ROP2基因的克隆及原核表达[EB/OL].(2006-01-18)[2025-08-05].http://www.paper.edu.cn/releasepaper/content/200601-217.点此复制
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