羊白介素12受体β2链的表达与纯化  

Expression and purification of ovine interleukin-12receptor β2 chain

作　　者：蒙学莲[1] 朱学亮[1] 张志东[1] MENG Xue-lian;ZHU Xue-liang;ZHANG Zhi-dong(State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃兰州730046

出　　处：《中国兽医学报》2020年第3期575-580,587,共7页Chinese Journal of Veterinary Science

基　　金：国家“十三五”重点研发计划资助项目(2016YFD0500108)。

摘　　要：利用PCR和基因合成获得了野生型羊IL-12Rβ2和密码子优化的IL-12Rβ2-A基因,亚克隆至pET30a(+),SDS-PAGE对这2个重组载体的表达结果进行比较,并优化了pET30a-12Rβ2-A的最佳表达条件;通过Western blot对镍琼脂糖凝胶纯化的融合蛋白His-12Rβ2进行了检测。结果显示,pET30a-12Rβ2-A的表达量明显高于pET30a-12Rβ2的表达;在筛选的pET30a-12Rβ2-A最佳表达和纯化条件下,成功获得了高纯度的融合蛋白His-12Rβ2,达到了预期的目标,为后续的功能研究奠定了基础。Ovine β2 chain of the interleukin(IL)-12 receptor(IL-12Rβ2)acts not only to as transducer of IL-12 signals but also as a gatekeeper gene and high affinity converter.At present,there are no data about the recombinant expression and purification of ovine IL-12Rβ2.This study was designed to optimize the expression condition and purification of the extracellular region of ovine IL-12Rβ2.The wild type(Wt)IL-12Rβ2 and codon optimized IL-12Rβ2-A were obtained by PCR and artificial gene synthesis,and directionally subcloned into prokaryotic expression vector pET30a(+),respectively.The expression conditions and expressed products were analyzed by SDS-PAGE.The expression condition of IL-12Rβ2-A gene was optimized and the fusion protein His-12Rβ2purified by Ni+affinity was detected by western blotting.The results showed that the expression level of pET30a-12Rβ2-A was significantly higher than that of pET30a-12Rβ2.Under the optimized expression and purification conditions,the highly purified fusion protein His-12Rβ2 was obtained successfully.The results indicated that the expected objective was achieved and laid a foundation for further functional research on IL-12Rβ2.

关 键 词：白细胞介素-12受体β2链 表达 纯化 优化 

分 类 号：S852.2[农业科学—基础兽医学]