ephrinB2真核表达载体的构建及其在Hela细胞的表达

目的构建ephrinB2重组真核表达载体,研究该质粒在细胞中的表达,为研究ephrinB2对肿瘤生长及血管生成的影响奠定基础。方法逆转录获得人ephrinB2全长cDNA序列,连接到pEGFPN1上,转化大肠杆菌,脂质体转染Hela细胞。RT-PCR检测转录情况,WB鉴定目的蛋白表达。结果酶切和测序证实正确构建重组质粒,并在Hela细胞中表达。结论成功构建了pEGFPN1-ephrinB2真核表达载体,并在Hela细胞中有效表达,为进一步研究ephrinB2的功能奠定基础。

Construction and in Vitro Expression of an Eukaryotic Vector Encoding ephrinB2

Objective To construct an eukaryotic plasmid expressing ephrinB2 and test whether the plasmid can be expressed in the eukaryotic cells in vitro for study the effect of ephrinB2 on tumor growth and angiogenesis.Methods The full-length ephrinB2 cDNA fragment were amplified by RT-PCR.Then the PCR product was inserted into eukaryotic fluorescent expressing vector p EGFPN1 by DNA recombinant technology and further transfected into HeLa cervical carcinoma cells.Expression of ephrinB2 in transfectants was examined by the fluorescence microscope,RT-PCR and Western blot.Results The 1kbp DNA fragment of ephrinB2 was correctly amplified.The recombinant pEGFPN1-ephrinB2 was successfully constructed and could be correctly expressed in the Hela cells.Conclusion This recombinant plasmid pEGFPN1-ephrinB2 and the resultant Hela cell model provide a basis for further study on the function of ephrinB2.