肿瘤抑制基因DOC-2真核表达载体的构建及在卵巢癌细胞HO-8910中的表达鉴定

Construction of The Eukaryotic Expression Vector pCDNA3.1-p93 and Its Expression in Ovarian Cancer Cell Line HO-8910

摘要: 目的构建肿瘤抑制基因DOC-2的真核表达载体pCDNA3.1-p93,将其转入人卵巢癌细胞系HO-8910中,对其基因表达进行相关检测,为进一步研究DOC-2的功能奠定基础。方法利用XhoI酶切含有p93cDNA的质粒得到p93cDNA基因片段,将其连接入真核表达载体pCDNA3.1,并通过酶切鉴定。用脂质体介导法将真核表达载体pCDNA3.1-p93转染HO-8910,通过G418筛选稳定表达克隆;用免疫组化法观察人DOC-2蛋白在HO-8910中的表达。结果构建了DOC-2的真核表达载体pCDNA3.1-p93,并通过酶切鉴定。免疫细胞化学结果显示pIRES2-EGFP-p93成功转入HO-8910中。结论成功构建了真核表达载体pCDNA3.1-p93并在HO-8910中得到稳定表达,为研究DOC-2蛋白的功能奠定了基础。

Abstract: Objective To construct an recombinant vector consisted of pCDNA3.1 with DOC-2 cDNA(p93) and transfect it into human ovarian cancer cell line HO-8910 in order to investigate the function of DOC-2 gene on ovarian cancer cell. Methods The recombinant vector (pCDNA3.1-p93) was constructed by XhoI and its correction was confirmed by restriction pattern. Then pCDNA3.1-p93 was transfected into HO-8910 by Lipofectamin and the positive clone 8910-p93 and 8910-pCDNA3.1 were selected by G418. Finally, the expression of...