丙酮酸脱氢酶复合物X蛋白的原核表达与血清学鉴定

目的获得丙酮酸脱氢酶复合物的X蛋白(Pro-X)的原核表达蛋白.方法采用RT-PCR技术从人淋巴细胞RNA中扩增出Pro-X的基因片段,克隆至pET28a( )表达载体进行诱导表达,并对表达产物进行Western blot和ELISA鉴定.结果成功构建了表达载体pET28a( )/Pro-X;表达产物能特异性的被原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)患者血清中的抗线粒体抗体识别.结论获得Pro-X蛋白的原核高效表达,为利用原核表达的丙酮酸复合物对PBC患者进行血清学检测进一步奠定了基础.

Prokaryotic expression and serum identification of X protein of dihydrolipoamide acetyltransferase compounds

Objective To acquire the X protein of dihydrolipoamide acetyltransferase gene (Pro-X) through prokaryotic expression. Methods The Pro-X gene was amplified from human lymphocytes with RT-PCR method, and was cloned into pET28a( ) vector to induce the Pro-X expression. The expressed product was evaluated by Western blotting and enzyme linked immunoabsordent assay. Results The expression vector pET28a( )/Pro-X was successfully constructed. The expressed product could be identified by the specific self-antibodies in the serum of primary biliary cirrhosis (PBC) patients. Conclusion High efficient expression of Pro-X lays the foundation for serum assay of PBC patient by prokaryotic expression of dihydrolipoamide acetyltransferase compounds .