cdx2真核表达质粒的构建

目的：构建含有cdx2全长基因的真核表达质粒并在人胃上皮细胞系中表达。方法：从人结肠上皮组织中提取总RNA,RT-PCR获得全长人cdx2基因的cDNA,测序证实序列正确后亚克隆入pcDNA3.1来构建重组真核表达载体pcDNA3.1/cdx2。使用脂质体将该重组质粒转染入人胃上皮细胞GES-1中,RT-PCR证明该细胞中出现人cdx2基因的表达。结果：酶切鉴定和测序显示靶基因cdx2克隆到pcDNA3.1质粒中,RT-PCR证实转染的人胃上皮细胞中有人cdx2基因的表达。结论：成功构建含有人cdx2基因的真核表达质粒。并在人胃上皮细胞中表达。

Construction of Eukaryotic Expression Plasmid Containing cdx2 Gene

Objective：To construct eukaryotic expression plasmid containing cdx2 full-length gene and express it in human gastric cell line.Methods： The total RNA was extracted from human colonic epithelium.The cDNA encoding human full-length cdx2 gene was obtained by RT-PCR.After the sequencing was confirmed by sequencing,the gene was subcloned to pcDNA3.1 to construct recombinant eukaryotic expression vector pcDNA3.1/cdx2.The recombinant plasmid was transfected into human gastric cell GES-1 by lipofectamine.Then the expression of human cdx2 gene in transfected cells was confirmed with RT-PCR.Results： Enzyme digestion analysis and sequencing showed that the target gene cdx2 was cloned into pcDNA3.1 plasmid.Expression of human cdx2 gene in the transfected human gastric cell was identified with RT-PCR.Conclusions ：The eukaryotic expression plasmid containing human cdx2 gene is successfully constructed and then expressed in human gastric cell.