Characterization of Expression of purH gene in Beijing Fatty Chicken ( Gallus gallus)

The specific expression of purH gene in eight different tissues of Beijing fatty chicken was investigated by HT-PCR.The full-length cDNA of purH was inserted into pEGFP-N3 multi-cloning sites to construct recombinant vector pEGFP-purH.The lipofectin method was used to transfect the pEGFP-purH into fibroblast cells.Bioinformatic analysis be transfected showed CDS region of the Beijing fatty chicken purH gene was 1 782 bp,which encoded a 483-aminoacids -long peptide.The recombinant pEGFP-purH was constructed with GFP as reporter gene to transfected into Beijing fatty chicken fibroblasts.The expression efficiency ranged from 10.3 % to 53.2 % in 24,48,and 72 h after transformation. RT-PCR and Western blotting analyses showed that pECFP-purH had been integrated into the genome of Beijing chicken fibroblast cell with normal expression level.In optimized condition,there was no significant effect (P0.05) on apoptosis ratio,growth and reduplication state comparing with the control group.