ABSTRACT
The specific expression of alpha1-AGP gene in eight different tissues of Beijing fatty chicken was investigated by RT-PCR. The full-length cDNA of alpha1-AGP was inserted into pEGFP-C1 multi-cloning sites to construct recombinant eukaryotic expression vector pEGFP-alpha1-AGP. The lipofectin method was used to transfect the pEGFP-alpha1-AGP into Beijing fatty chicken fibroblast cells. The open reading frame of Beijing fatty chicken alpha1-AGP gene was 612 base pairs in length, which was expressed higher in liver and lung than in muscle. This gene did not express in heart and kidney. The expression efficiency ranged from 31.3% to 47.6% in 24, 48, and 72 h after transformation. The green fluorescence mainly concentrated in the nucleus. With the increase of the expression of green fluorescence, granula was observed in the nucleus. RT-PCR and Western blotting analyses showed that pEGFP-alpha1-AGP had been integrated into the genome of Beijing chicken fibroblast cell with normal expression level. In optimized condition, there was no significant effect (P>0.05) on apoptosis ratio, positive cell shape, growth and reduplication state comparing with the control group. This research established the foundation for further function research of alpha1-AGP gene and application in transgenic animal cloning.
