【摘要】 目的:将骨保护素(osteoprotegerin,OPG)与人血清白蛋白(human serum albumin,HSA)基因融合,以构建高效分泌表达的巴斯德毕赤酵母菌株。方法:通过RT-PCR扩增OPG基因,构建pHILD2-HSA-OPG质粒,并对HSA-OPG分泌表达产物进行了ELISA检测。结果:pHILD2-HSA-OPG表达质粒经鉴定与测序证明正确,并在毕赤酵母GS115中能够实现较高表达。结论:成功构建了高效分泌表达重组骨保护素融合蛋白(rhOPG-HSA)的毕赤酵母菌株,为进一步研究其生物学活性奠定了基础。更多还原

【Abstract】 Objective:To fuse the gene of osteoprotegerin(OPG) with that of human serum albumin(HSA) in order to construct a Pichia pastoris yeast strain with high-efficient expression of HSA-OPG.Methods:The gene of OPG was amplified by RT-PCR and the plasmid of pHILD2-HSA-OPG was constructed.HSA-OPG fusion protein was detected with ELISA kit.Results:The expression plasmid of pHIL-D2-HSA-OPG was proved to be correct after plamid identification and sequencing,and rhHSA-OPG was relatively efficiently expressed in Pichia pastoris.Conclusion:Pichi pastoris yeast strain with high-efficient expression of HSA-OPG is successfully constructed,providing a basis for further study of its bioactivity.更多还原