中国畜牧兽医2016，Vol.43Issue(12)：3121-3126,6.DOI:10.16431/j.cnki.1671-7236.2016.12.006
大约克猪 SLA-1原核表达载体的构建及表达
Construction and Expression of SLA-1 Prokaryotic Expressing Vector in Yorkshire Pig
摘要
Abstract
In order to construct the prokaryotic expressing vector of SLA-1 derived form York-shire pig and express the interest of protein,a pair of primers was designed to amplify the extra-cellular domain of SLA-1 gene from Yorkshire pig (named SLA-1-DYKe)by PCR.Then the PCR product was cloned into pMD® 1 9-T Simple Vector and transformed into Escherichia coli TOP10. After cleaved by Nde Ⅰ and Xho Ⅰ,the positive clones were selected to be sequenced.Analy-zing by biological soft,the fragment from positive clone with correct sequence was inserted into pET-28a (+)and transformed into E .coli BL21 (DE3).After induction and expression,the in-terest of protein was detected by SDS-PAGE.The results showed that the extracellular domain of SLA-1-DYKe was successfully amplified with the fragment length of 837 bp.The interest of SLA-1 gene was successfully cloned into pMD® 1 9-T Simple Vector and the positive recombinant plasmids with correct sequences were obtained.The SLA-1-DYKe from positive recombinant plasmids was further inserted into pET-28a(+).After transformed into E .coli BL21(DE3)and induction,the SLA-1-DYKe was successfully expressed.The molecular weight of the protein was about 34 ku.It was concluded that the prokaryotic expressing vector of SLA-1 was constructed successfully from Yorkshire pigs and then the expressed protein was obtained,which would lay a base for studying on the structure and function of SLA-1 from Yorkshire pig in the future.关键词
大约克猪/SLA-1 基因/原核表达载体Key words
Yorkshire pig/SLA-1 gene/prokaryotic expressing vector分类
生物科学引用本文复制引用
张秀娟,杨杰,孙永强,高凤山..大约克猪 SLA-1原核表达载体的构建及表达[J].中国畜牧兽医,2016,43(12):3121-3126,6.基金项目
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(31172304) （31172304）
2015年辽宁省大学生创新创业训练计划项目:育-大约克猪 SLA-1原核表达载体的构建及表达研究(20150086) （20150086）
2014年大连大学大学生创新创业训练计划项目重点项目:育-大约克猪 SLA-1原核表达载体的构建及表达研究(2014066) （2014066）
