抗凋亡基因bcl-2 cDNA表达型质粒的构建及序列分析  

The construction and sequencing of the expression plasmid of the apoptosis-resistent gene bcl-2

作　　者：何庆[1] 苏京[2] 刘铭[3] 王保平[2] 王晶[2] 陈克勤[2] 尹潍[1] 

机构地区：[1]天津医科大学总医院内分泌科,天津300052 [2]诺和诺德中国研究发展中心,北京100176 [3]天津医科大学老年病研究室

出　　处：《天津医科大学学报》2004年第1期14-17,共4页Journal of Tianjin Medical University

基　　金：天津市自然科学基金资助课题 (编号 :983702411)

摘　　要：目的 :克隆抗凋亡基因bcl-2,并构建其表达型质粒 ;进行序列分析 ,为基因转染作准备。方法 :应用PCR定向克隆技术 ,克隆SD大鼠鼠脑bcl-2全编码cDNA序列 ;应用双脱氧末端终止法进行重组质粒中bcl-2的cDNA序列分析 ;利用基因重组技术 ,将bcl-2全编码cDNA序列 ,导入pCMV -Tag -1表达型质粒。结果 :成功地克隆SD大鼠bcl-2全编码cDNA序列 ,并将其成功地导入pCMV -Tag -1表达型质粒 ;经序列分析 ,发现bcl-2全编码序列与GeneBank中的相应序列相比 ,有3个碱基出现差别 ,分别为171位a→g,233位a→c ,530位 g→a,均为同义突变。结论Objective:Cloning the full length cDNA of bcl-2gene,which believed to be has the effect of resisting the apoptosis,and constructing the expression plasmid;sequencing the bcl-2gene,to prepare for gene transfection.Methods:Cloning the full length cDNA of bcl-2gene of SD rat brain tissue by PCR target cloning method;Sequencing the full length cDNA of bcl-2gene by dideoxy-nucleotide chain termination method;Putting the the full length cDNA of bcl-2gene into expression plasmid pCMV-Tag-1by gene recombination method.Results:Full length cDNA of bcl-2gene was cloned and constructed into the expresˉsion plasmid of pCMV-Tag1successfully.The cDNA sequence showed that only three bases were different from that in Gene Bank but the difference was believed to be synonymous muˉtation.Conclusion:This work provided the base for the processing study that was on the transfection of the anti-apoptosis bcl-2gene to the islet cell.

关 键 词：bcI-2基因 克隆 逆转录聚合酶链反应 表达型质粒 序列分析 

分 类 号：Q785[生物学—分子生物学]