人源核不均一核糖核蛋白E1真核表达载体在神经细胞中的表达

Expression of recombinant plasmid of homo heterogeneous nuclear ribonucleoprotein E1 in SH-SY5Y cells

背景:人源核不均一核糖核蛋白E1功能广泛,可参与神经系统骨架蛋白的表达。目的:为深入研究其在神经细胞中的作用,构建其真核表达载体,观察其在神经细胞中的表达。方法:利用真核表达载体pcDNATM4/His C,通过亚克隆构建核不均一核糖核蛋白E1-pcDNATM4/His C重组质粒,经酶切、测序鉴定,通过转染神经细胞SH-SY5Y,采用western-blot,RT-PCR鉴定核不均一核糖核蛋白E1重组质粒的表达,并观察转染细胞的生长现象。结果与结论:成功构建了核不均一核糖核蛋白E1的真核表达载体,mRNA和蛋白水平上均证实了该质粒可在神经细胞SH-SY5Y中正确表达。SH-SY5Y细胞在转染核不均一核糖核蛋白E1后表现为加速生长。提示该蛋白对神经细胞的生长发育具有重要的作用。该载体为进一步研究核不均一核糖核蛋白E1在神经系统中的功能提供了前提条件。

BACKGROUND：The functions of homo heterogeneous ribonucleoprotein E1 are very wide. It can participate in the expression of skeleton proteins in the nervous system. OBJECTIVE：To construct the recombinant plasmid of homo heterogeneous ribonucleoprotein E1 and observe its expression in nerve cells for further studying the functions of it in neurocytes. METHODS：Using pcDNATM4/His C, the homo heterogeneous ribonucleoprotein E1 was subcloned into recombinant plasmid E1-pcDNATM 4/His C, fol owed by enzyming and sequencing. After SH-SY5Y cells were transfected with the recombinant plasmid, western blot analysis and real time RT-PCR were used to detect the expression of homo heterogeneous ribonucleoprotein E1 in SH-SY5Y cells. And the growth of SH-SY5Y cells was observed. RESULTS AND CONCLUSION：We successful y constructed the eukaryotic expressed vector of homo heterogeneous ribonucleoprotein E1. The recombinant plasmids were verified to express in SH-SY5Y cells correctly at mRNA and protein levels. And SH-SY5Y cells generated quickly after homo heterogeneous ribonucleoprotein E1 was over-expressed. The homo heterogeneous ribonucleoprotein E1 is an important protein in neural development. And this vector offers the premise for further studying its function in nervous system.