上皮细胞剪接调节因子-1真核表达载体的构建及表达Construction and expression of the eukaryotic expression vector of human epithelial splicing regulatory protein 1
李竹;孟祥玉;周晶琳;高晓虹;陈志鸿;
摘要(Abstract)：
目的构建人内皮细胞剪接因子-1真核表达载体pcDEF3-ESRP1,并观察其在HeLa细胞的表达。方法以HeLa细胞cDNA为模板,用RT-PCR方法扩增ESRP1全编码区基因片段,将其连接入真核表达载体pcDEF3,并通过酶切及测序进行鉴定;通过脂质体介导法将真核表达载体pcDEF3-ESRP1转染至HeLa细胞,Western blot检测外源性ESRP1蛋白在HeLa细胞中的表达。结果 RT-PCR扩增得到人ESRP1 cDNA片段,重组质粒经酶切鉴定和DNA序列分析确定得到真核表达载体pcDEF3-ESRP1;Western blot实验证实转染重组质粒的HeLa细胞存在外源性的人ESRP1表达。结论成功构建人ESRP1真核表达载体pcDEF3-ESRP1,为进一步研究ESRP1的生物学功能奠定了实验基础。
关键词(KeyWords)： 上皮细胞剪接调节因子-1;真核表达载体;脂质体
基金项目(Foundation): 黑龙江省自然科学基金(LC2016040)
作者(Authors): 李竹;孟祥玉;周晶琳;高晓虹;陈志鸿;
DOI: 10.13799/j.cnki.mdjyxyxb.2017.02.005
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