高迁移率蛋白A2基因真核表达载体构建与表达鉴定

目的：构建高迁移率蛋白A2（HMGA2）基因的真核表达载体，观察其在真核细胞中的表达，为进一步研究HMGA2 基因功能奠定理论基础。方法：采用RT-PCR 方法从人乳腺上皮HBL-100 细胞中扩增HMGA2 基因全长cDNA，扩增产物经双酶切及凝胶回收后，直接克隆至黄绿色荧光真核表达载体YFP-C1中，构建YFP- C1-HMGA2重组质粒。将YFP-C1-HMGA2真核表达载体转染到人前列腺癌PC3细胞中，通过RT-PCR方法验证所构建的真核表达质粒YFP-C1-HMGA2的正确性。结果：RT-PCR获得长度为351　bp的HMGA2全长cDNA，经YFP-C1真核表达载体克隆、酶切鉴定及测序分析，测序结果示基因序列与GenBank序列一致，证实真核表达质粒YFP-C1-HMGA2构建成功，并且能够在真核细胞中表达。结论：成功克隆了HMGA2基因，构建了YFP-C1-HMGA2重组质粒。

Construction and expression identification of eukaryotic expression vector of HMGA2 gene

Objective To construct the recombinant eukaryotic expression vector of HMGA2 gene and observe its expression in eukaryotic cells,and laid a theoretical foundation for further study of HMGA2 gene function.Methods HMGA2 gene full-length cDNA from human breast epithelial cell HBL-100 was amplified by RT-PCR;after double-digestion and gal recovery HMGA2 gene was cloned into YFP-C1 eukaryotic expression vector.After eukaryotic expression vector YFP-C1-HMGA2 was transfected into human prostate cancer cells,and th...