禽白血病病毒p27基因在毕赤酵母中的表达

Expression of Avian Leukosis Virus p27 Gene in Pichia pastoris

为获得禽白血病病毒p27蛋白,利用DNA重组技术,将扩增的p27基因亚克隆至酵母表达载体pPIC9K中,并将制备的酵母表达质粒电转至毕赤酵母细胞GS115中,通过SDS-PAGE和Western blot技术检测p27蛋白在不同条件下的表达情况,同时对诱导表达条件进行系统的优化。结果表明,成功构建出pPIC9K-p27酵母表达质粒,当诱导时长为24 h,甲醇诱导浓度为1%,BMMY培养基的pH值为7.0,诱导温度为28℃时,目标蛋白rp27的表达量最高可达26.2μg/mL。本研究为进一步开发禽白血病病毒检测试剂盒奠定了基础。

In order to obtain p27 protein of Avian leukosis virus,the p27 gene was cloned into yeast expression vector pPIC9K to obtain a yeast expression plasmid that was electroporated into Pichia pastoris cell GS115 using DNA recombination technology.The expression of p27 protein was optimized under different conditions and the expressed p27 protein was analyzed by SDS-PAGE and Western blot.The results showed that the yeast plasmid pPIC9K-p27 was successfully constructed and the optical p27 protein at 26.2μg/mL was expressed with 24 h induction,1%methanol,BMMY medium at pH7.0 and the temperature at 28°C.The availability of the p27 protein provided the basis for further development of Avian leukosis virus detection kit.