Construction and expression of recombinant plasmids encoding chicken UAF1 gene

To construct the prokaryotic expression vector of chicken ubiquitin-related factor 1(chUAF1),total RNA was isolated from chicken bursa of fabricus and then cDNA was synthesized by RT-PCR.According to the features of the mammalian genome which containning high GC content in 5′-end and low GC content in 3′-end,we designed two pairs of primers to amplified two fragments of chUAF1 by PCR.The full length of chUAF1 gene was reconstitued by linking a same restriction site in the overlap of two fragments and then chUAF1 gene was cloned into pMD18-T vector for sequencing.The correct chUAF1 gene was subcloned into pET28a vector to construct the recombinant plasmids of pET28a-chUAF1.The recombinant plasmids were tansformed into BL21(DE3) competent cells.Expression of chUAF1 was induced by IPTG and identified by SDS-PAGE.The results showed chUAF1 was successfully expressed in E.coli.expression system.