逆转录PCR克隆L－选择素cDNA  

Amplifying and cloning of human L─selectin cDNA by reverse transcription and PCR method

作　　者：赵智晖[1] 赵克森[1] 汲言山[1] 沈倍奋[1] 马宏进[1] 

机构地区：[1]第一军医大学病理生理教研室,军事医学科学院基础所

出　　处：《中国病理生理杂志》1996年第1期26-29,共4页Chinese Journal of Pathophysiology

摘　　要：本文为从转录水平研究Ｌ─选择素表达的调节机制，利用Ｇｏｌｄｋｅｙ软件及ＥＭＢＬ数据库选出人与大鼠Ｌ─选择素高同源性但与其它细胞因子及其受体无高同源性的ｃＤＮＡ片段，设计ＰＣＲ引物，一步法分离Ｂ系Ｒａｊｉ细胞ＲＮＡ，逆转录ＰＣＲ扩增目的片段，ＨｉｎｆⅠ酶切初步确定后，将片段插入ＰＵＣ１９质粒ＨｉｎｃⅡ位点，转化ＪＭ１０９大肠杆菌，经测序证实扩增克隆片段与设计相符。In order to investigate the mechanism of L─selectin expression attranscription level,human L─selectin cDNA sequence was compared with that of therat,high homologic segment between these two but low homology with other cytokinesand receptors was extracted,and PCR (polymerase chain reaction)primers weredes igned by using Goldkey ver.1.0.With single─step method,Raji cell total RNAwas isolated and its reverse transc ript were used as PCR templets.567bp segmentswere amplified.After identification by Hinc Ⅱ digestion,the amplified segment wascloned to Hinc Ⅱ site of PUC 19 vector,and then the recombinant plasmids weretransferred to JM 109 E.Coli.The inserted segment was sequenced and proved to beidentical with published sequence.

关 键 词：逆转录酶类 克隆 选择素 CDNA 聚合酶链反应 

分 类 号：R392.32[医药卫生—免疫学]