Study On The Cloning And Expression Of α-IFN Gene In Beijing Fatty Chicken

In this paper, we made the total RNA of liver from Beijing Fatty Chicken as a template, cloned target genes to PGEM-T Easy through RT-PCR and constructed Beijing Fatty Chicken's cloning vector of PGEM-T-α–IFN. constructed Beijing Fatty Chicken expression vector pEGFP-T-α–IFN through the method of double restriction enzyme, and expression in target cells. The results as follows:1. we made the total RNA of liver from Beijing Fatty Chicken as a template, to clone chickenα-IFN gene through RT-PCR method, a size of 582 bp, and the gene sequences submitted to GenBank.2. we made fibroblast cells of Mongolian horse and aletai sheep cultivated by direct adherent culture methods as target cells, and the living rate of cells, the growth curve, microbial contamination, karyotype, with the Spectrum of enzymes and green fluorescent protein from sources such as identification of gene expression, the target cell growth has been proved healthy and good shape.3. We construct pEGFP-C1-α-IFN eukaryotic expression vector of Beijing Fatty Chicken using SalI, EcoRI double digestion. The result of expression in target cells showed that: the rate of transfection in Mongolian horse is up to 39.42 percent after 72h; aletai sheep is up to 40.21 percent after 48 hours. A higher rate of Green fluorescent indicated a dispersion ofα-IFN in the target cells.