水貂阿留申病病毒VP2截短基因的原核表达

Prokaryotic Expression of VP2 Truncated Gene of Aleutian Mink Disease Virus

为获得水貂阿留申病病毒G株(ADV-G株)VP2截短基因表达的蛋白,将ADV-G株VP2基因的主要抗原区片段克隆至原核表达载体pGEX-4T-1中,通过不同浓度的IPTG,不同诱导时间以及加入不同的化学分子伴侣(甘油、葡萄糖、蔗糖、二甲基亚砜、乙醇)进行诱导表达,确定目的蛋白表达的最佳条件。经SDS-PAGE和Western blot分析,表明终浓度为1mmol/L的IPTG诱导6h目的蛋白的表达量最高,分子质量约为53ku。SDS-PAGE分析表明,葡萄糖和乙醇抑制了目的蛋白的表达,而蔗糖、甘油和二甲基亚砜提高了目的蛋白的表达量,获得的目的蛋白具有良好的反应原性。

This experiment was conducted to obtain the protein expressed by VP2 truncated gene of Aleutian mink disease virus and study its reactionogenicity.The fragment with main antigenic epitope of VP2 gene of Aleutian mink disease virus（ADV-G）was cloned into a prokaryotic expression plasmid pGEX-4T-1.Subsequently,different concentrations of IPTG were used to induce the target protein at different induction time.At the same time,different chemical molecular chaperones were applied to make sure the optimum expression condition.The specificity of the expressed protein was identified through SDS-PAGE and Western blot.The results showed that the target protein was about 53 ku in size and it reached its peak after 6hexpression with 1mmol/L IPTG.The SDS-PAGE results also showed that glucose and ethanol had no influence on the expression,while sucrose,glycerin and dimethyl sulfoxide could increase the expression of target protein.The target protein had good reactionogenicity,which laid a foundation for the diagnosis and prevention of Aleutian disease.