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目的基因工程重组表达2-氧戊二酸脱氢酶复合体E2(OGDC-E2)融合蛋白。方法用PCR方法扩增得到OGDC-E2的基因片段,插入表达载体pET28a(+),构建重组表达载体pET28a(+)-OGDC-E2,测序正确后在大肠杆菌BL21(DE3)中表达。结果获得OGDC-E2融合蛋白,经SDS-PAGE分析,相对分子质量与预期大小一致;经免疫学鉴定,重组抗原片段具有抗线粒体抗体二亚型(AMA-M2)的免疫原性。结论基因重组表达的融合蛋白将有利于原发性胆汁性肝硬化(PBC)的实验诊断。
Objective To genetically engineer the 2-oxoglutarate dehydrogenase complex E2 (OGDC-E2) fusion protein. Methods The gene fragment of OGDC-E2 was amplified by PCR and inserted into the expression vector pET28a (+). The recombinant plasmid pET28a (+) - OGDC-E2 was constructed and expressed in E. coli BL21 (DE3) after sequencing. Results The OGDC-E2 fusion protein was obtained. SDS-PAGE analysis showed that the molecular weight of OGDC-E2 was consistent with the expected size. The immunogenicity of recombinant OGDC-E2 fusion protein was confirmed by immunohistochemistry. Conclusion The recombinant fusion protein will be helpful for the experimental diagnosis of primary biliary cirrhosis (PBC).