人NKG2ACD94的基因克隆和真核共表达载体的构建-检验医学.PDF

·159· 文章编号：1673-8640(2007)02-0159-04中图分类号：Q503 文献标识码：A 人NKG2A、CD94的基因克隆和真核共表达载体的构建 焦玉莲，王来城， 张捷， 崔彬， 马春燕，赵跃然 (山东大学山东省立医院科研中心，山东济南250021) 摘要：目的构建真核共表达载体pNKG2A-IRES—CD94。方法提取人外周血淋巴细胞总RNA，逆转录．聚 I和sal 分析。pMD-CD94经XbaI酶切，插入相应酶切的真核表达载体plRES(多克隆位点B)，NKG2A经Xho I和Mlu 达质粒pNKG2A·IRES—CD94，为研究NKG2A和CD94基因的功能奠定了基础。 关键词：NKG2A；CD94；共表达质粒；内部核糖体进入位点 Constructionofanovel vector CO-expression JIAO system：pNKG2A-IRES-CD94Yulian，WANGLaicheng， Yueran． Research Province ZHANG施，CUIBin，MA (MedicalCentre，$handong Chunyan，ZHAO Hospital，Shandong J／nan University，Shandong250021，China) Toconstructa is Abstract：OMecttveeukaryotic CO—expressionplasmid，pNKG2A—IRES-CD94，whichcapable ofstable of am— NKG2AandCD94 ThehumanNKG2AandCD94 were CO-expression gene．Methods genefragments reverse chain thetotalRNAisolatedfromhuman plifiedby transcriptionpolymerasereaction(RT-PCR)from peripherM blood andclonedintothe vectortoconstruct and were lymphocyte pMDl8-T pMD—NKG2ApMD-CD94repectively，which identified Sal was withXbaI and I andinsertedinto bysequenceanalysis．TheplasmidpMD—CD94digested eukary- otic vector toobtainrecombinanthumanCD94 CO-expressionplRES expressionplasmidplRES—CD9