摘要
设计1对特异性引物,采用PCR方法从pMD18-TCHIFN-γ重组载体中扩增得到广西三黄鸡γ-干扰素基因,然后将该基因克隆入含有绿色荧光报告基团EGFP的真核表达载体pEGFP-N1中,构建成pEGFP-N1CHIFN-γ重组质粒,PCR鉴定结果和测序结果均显示,γ-干扰素基因正确地插入到真核表达载体中。使用脂质体转染法将pEGFP-N1CHIFN-γ转染Vero细胞,荧光显微镜下成功地观察到绿色荧光,说明γ-干扰素基因成功表达。
One pair of specific primers was designed to clone Guangxi Sanhuang chicken interferen-γgene from the recombinant vector pMD18-CHIFN-γ. And then the interferen-γgene was inserted into eukaryotic expression vector pEGFP-N1 which included one EGFP fluorescence gene. The recombinant vecter pEGFP-N1 CHIFN-γ was proved correct by PCR and sequencing,then co-transfected into Vere cells,The fluorescence was detected by fluorescopy,so it was proved that the interferon-γwas expressed successfully.
出处
《西南农业学报》
CSCD
2008年第1期167-169,共3页
Southwest China Journal of Agricultural Sciences
基金
广西留学基金项目(桂科回0542006)
关键词
鸡γ-干扰素
绿色荧光
真核表达
chicken intefferen-γ
green fluorescence
eukaryotic expression
