膜联蛋白V真核表达载体的构建

CONSTRUCTION OF EUKARYOTIC EXPRESSION VECTOR CONTAINING HUMAN ANNEXIN V GENE

目的 :构建人膜联蛋白 V基因真核表达载体 ,探讨人膜联蛋白 V的生理功能。方法 :PCR方法扩增含 Eco RI及 Bam HI酶切位点的膜联蛋白 V基因序列 ,对真核表达载体 pc DNA 3.1 (- )和膜联蛋白 V基因扩增产物进行双酶切 ,用连接酶将二者连接并转化到大肠杆菌 BL 2 1 ,重组质粒经测序鉴定 ,称为 pc DNA3.1 (- ) Ax V。结果 :酶切琼脂糖电泳分析 pc DNA3.1 (- )Ax V中含有膜联蛋白 V基因 ,序列分析表明与文献报道的序列完全一致。结论 :成功地构建了人膜联蛋白 V基因的真核表达载体 。

Objective:To construct eukaryotic expression plasmid of human annexin V gene for further study.Methods:Annexin V gene containing EcoRI and BamHI endoenzyme site was obtained by using PCR.Double enzyme digestion was conducted for pcDNA3 1(-) and PCR product of annexin V gene.Both fragements were connected by using ligase and transferred to BL21.Reconstituted plasmid was examined by auto sequenceing assay and was namedas pcDNA3.1(-)AxV.Result:The pcDNA3 1(-)AxV plasmid was confirmed to contain correct nucleotide sequence for human annexin V gene by restriction endonuclease digestion and sequence analysis.Conclusion:A eukaryotic expression plasmid containing human annexin V gene was successfully constructed and was helpful for the research for the function of annexin V.