人PSF真核表达载体构建及其在CHO细胞中的表达

目的是构建人PSF基因真核表达载体pEGFP-N1-PSF,并在检测其在CHO细胞株中的表达情况。应用DNA重组技术和PCR方法从人宫颈癌Hela细胞中扩增PSF基因,插入pEGFP-N1真核表达载体,构建重组质粒pEGFP-N1-PSF并测序鉴定。将pEGFP-N1-PSF瞬时转染CHO细胞,通过Western blot和RT-PCR方法检测PSF的表达,荧光显微镜下观察绿色荧光蛋白表达。结果 CHO细胞转染pEGFP-N1-PSF真核表达载体后,RT-PCR和Western blot实验发现,在RNA和蛋白水平有PSF的表达,在荧光显微镜下可以观察到融合蛋白EGFP/PSF的表达。成功构建pEGFP-N1-PSF真核表达载体,证实其在CHO细胞中可以表达。

Construction of Eukaryotic and Identification of its Expression Vector of Human PSF Expression in CHO Cell Line

To construct eukaryotic expressing vector pEGFP-N1-PSF,and investigate its expression of RNA and protein in CHO cells,PSF gene was obtained by PCR and DNA recombination from Hela cell line.The products were cloned into pEGFP-N1.The recombined plasmid pEGFP-N1-PSF was sequenced and transfected into CHO cell.The expression of PSF was identified by RT-PCR and Western Blot.The expression of green fluorescent fusion protein was used to identify the effect of transfection.After plasmid pEGFP-N1-PSF was transfected into CHO,the expressed product of PSF was observed by Western blot and RT-PCR in protein and RNA level.The green fluorescent protein could be observed by fluorescence microscope after transfection.The recombinant eukaryotic expression vector of pEGFP-N1-PSF could be constructed and expressed in CHO cell line.