摘要
目的:为研究我国地方三品系猪SLA-1原核表达。方法:选取3个国内地方品系猪荷包、烟台黑猪及莱芜黑猪的SLA-1等位基因,PCR扩增地方品系猪SLA-1胞外区基因,再将胞外区基因克隆至pMD19-T Simple Vector,阳性重组子经过酶切,然后连接至pET-21a载体,构建其原核表达载体,并转化宿主菌BL21进行诱导表达。结果:PCR结果显示三个品系猪SLA-1胞外区大小约840bp,经克隆后成功连接至pMD19-T Simple Vector,经酶切回收后插入片段又成功连接至pET-21a载体,并在BL21成功表达,表达蛋白大小约31 kDa,与设计值大小相符。蛋白表达量约5%。结论:该研究成功表达了我国三个地方品系猪SLA-1胞外区,为下一步研究其空间结构及功能奠定基础。
Objective： To study the prokaryotic expression of SLA - 1 derived from three local breeds of swine. Method： SLA - 1 alleles from three local breeds of swine including Hebao, Yantai Black and Laiwu Black were selected to amplify the extracellular domains of SLA - 1. The extracellnlar domain of SLA - 1 was cloned into pMD 19 - T Simple Vector. The positive recombinant plasmids were cleaved and ligated into pET -21a to construct the prokaryotic expression vector and then they were transformed into Escherichia coli BI21 to be induced to expression. Result：The PCR results showed that the extracellular domains of SLA - 1 were about 840 bp and the PCR products were ligated into pMD 19 -T Simple Vector successfully. After cleavage and purification,the inserted fragments were ligated into pET 21a and expressed in Escherichia coli BL21 successfully. The expressed proteins were about 31 kDa,which was consistent with the theoreti- cal value and the content of the expressed protein was about 5%. Conclusion ：The extracellular domains of SLA - 1 genes from three local breeds of swine were expressed successfully in this study, which will lay the basis for studying their spatial structure and function in future.
出处
《生物技术》
CAS
CSCD
北大核心
2014年第1期16-19,共4页
Biotechnology
基金
国家自然科学基金项目("猪源病毒CTL多肽表位与SLA I结晶研究"
No.31172304)
2013年辽宁省大学生创新创业训练计划项目("地方三品系猪SLA-1原核表达载体构建及表达研究"
No.201311258042)
2012年大学生创新创业训练计划项目("地方三品系猪SLA-1原核表达载体构建及表达研究"
No.2012087)资助~~
关键词
猪
SLA-1
原核表达
载体
Swine
SLA - 1
Prokaryotic expression
Vector
