广西眼镜蛇神经毒素的原核表达

Prokaryotic expression of cobrotoxin from Naja naja atra Guangxi

利用拼接PCR法从广西眼镜蛇基因组DNA中获得CT成熟肽序列,序列分析显示,广西眼镜蛇与其他地区眼镜蛇的CT基因成熟肽序列具有较高的同源性,同时成功构建了pET30 a(+)-CT原核表达载体;研究结果表明,利用拼接PCR的方法能有效拼接两个外显子序列,广西眼镜蛇CT基因具有较高的保守性,CT基因可在大肠杆菌中表达。

The objective of current study was to obtain Naja naja atra Guangxi venom neurotoxin -- Cobrotoxin （CT） in vitro using splicing PCR. The result showed that CT gene mature peptide sequences was conserved among several species. And prokaryotie expression pET30 a （＋）-CT vector was successfully constructed. The current study suggested that using splicing PCR method could effectively spliced two exon sequences; Naja naja atra Guangxi CT gene had a high conservation, and could express in Escherichia coli.