中文摘要：

目的研究汉坦病毒76-118株包膜糖蛋白G2与β3整合素之间的相互作用。方法根据HV76—118株M基因和G3整合素基因序列设计引物，分别以质粒M56和Hela细胞cDNA为模板通过PCR和基因重组获得带有EGFP标签的G2和带有FLAG标签的β3整合素膜外区片段融合表达质粒，并且在HEK293细胞中进行表达。将表达成功的G2与β3整合素片段进行免疫共沉淀，用Western blot检测它们之间的相互作用。结果成功构建了G2和β3整合素膜外区各片段的融合表达质粒。将各表达质粒转染HEK293细胞进行瞬时表达，Western blot结果显示β3整合素膜外区各功能片段在细胞裂解上清中获得大量表达，而G2蛋白膜外区仅有81－140片段在细胞裂解上清中大量表达。将G2 81—140片段与β3整合素膜外区各功能片段进行免疫共沉淀的结果显示，仅G2 81—140片段和83整合素27—133片段在细胞中以复合物存在，提示两者之间可能有直接的相互作用。结论汉坦病毒76-118株包膜糖蛋白G2 81—140片段与β3整合素27—133片段在细胞中以复合物的形式存在，两者之间存在着相互作用，为β3整合素作为汉坦病毒受体提供了进一步的证据。

英文摘要：

To study the interaction of hantavirus envelope G2 protein with β3 intergrin protein in vivo, EGFP-tagged hantavirus envelope G2 fragments expression vectors and Flag-tagged β3 intergrin fragments expression vectors were constructed by PCR and gene recombination with the templates of plasmid M56 and eDNA from Hela cells. Transient expression of these recombinant proteins in HEK293 cells was detected by Western blot. Only G2 81-140 fragment and all β3 intergrin fragments were found in the supernatant of cell lysate. Pairs of the recombinant expression vectors were co-transfected into HEK293 cells and the interaction of hantavirus envelope G2 protein with β3 intergrin was identified by co-immunoprecipitation. The interaction between them could be identified by co-immunoprecipitation only in cells co-transfected with G2 81-140 and β3 27-133 fragments expression vectors. Our result shows that hantavirus envelope G2 protein and β3 intergrin may co-exist in the cells in the form of compound, namely there may be direct interaction between them. The interaction site may lie in G2 Nterminal 81--140 and β3 intergrin 27-133 amino acids.