高致病性禽流感H5N1病毒M1蛋白的重组表达及在感染检测中的初步应用

Expression of recombinant matrix protein 1 from H5N1 influenza virus and its application in diagnosis of infection

目的:原核表达重组高致病性禽流感基质蛋白M1并分析其在感染检测中的作用。方法:根据NCBI公布的基质蛋白M1的核苷酸序列设计特异性引物,用高致病禽流感H5N1病人分离的病毒提取病毒RNA进行反转录合成cD-NA,扩增基质蛋白M1基因。将M1基因克隆到pQE30原核表达载体进行非融合表达,纯化重组表达的蛋白并用免疫学方法检测分析。结果:克隆了高致病禽流感H5N1病毒基质蛋白M1基因,并采用pQE30原核表达载体成功表达该蛋白。以该重组蛋白为检测包被抗原建立间接ELISA,检测6份病人血清标本均为阳性,而正常对照人群为阴性。结论:高致病禽流感H5N1病毒M1蛋白可以应用于感染检测中。

Objective： To express M 1 of H5N1 influenza virus and analyze its function in in diagnosis of infection.Methods： The cDNA of an H5N1 influenza virus（2006/Shenzhen） and specific primers were used to amplify the M1 gene.After sequencing,M1 gene was subcloned into vector pQE30 to express M1 protein.Further, Purified M1 protein was analyzed by immunological method.Results： The pQE30-M1 expression vector was successfully constructed.Recombinant M1 protein was purified with Ni-NTA affinity chromatography and further used as detection antigen for indirect ELISA.All the 6 sera samples from patients were positive while samples from healthy donor were negative.Conclusion： The recombinant M1 protein can be applied in diagnosis of H5N1 infection.