海洋双RNA病毒VP2e基因的克隆和表达

Cloning and Expression of the VP2e Gene of Marine Birnavirus in E.coli

从海洋双RNA病毒(MABV)Y-6毒株基因组中克隆到VP2 e基因,将其连接到GST融合表达载体pGEX-6P-1,在大肠杆菌中表达,并对其表达条件进行了优化。通过温度和诱导物浓度优化试验,经SDS-PAGE和Western-Blot检测表明,在温度为37℃、IPTG浓度为0.1mmol/L时,表达的融合蛋白含量最高,占细菌总蛋白的28.6%。

The VP2e gene was amplified from the genome of marine birnavirus （MABV） by RT- PCR. Then, the PCR products were cloned into the GST fusion expression vector pGEX - 6P - 1 and expressed in E. coll. The conditions including expression temperature and IPTG - induction concentration were optimized. The results of SDS - PAGE and Western - blot showed that the expression level of the fusion protein occupies 28.6 % of the total bacterial proteins under the optimal conditions （37℃, 0. 1mmol/L IPTG）.