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目的 :构建人骨形态发生蛋白 -7(bonemorphgeneticprotein -7,BMP -7)巴斯德毕赤酵母(Pichiapastoris)表达重组子。方法 :以本室保存的重组质粒 pBV220/BMP -7为模板,经PCR扩增出人BMP -7成熟肽编码序列 ,先以A -T连接方式克隆入pGEM -Teasy载体,再将其克隆到毕赤酵母分泌型表达载体 pPIC9中,重组质粒线性化后PEG法转化入毕赤酵母菌株GS115,用PCR方法筛选转化子。结果 :DNA序列分析和PCR鉴定表明成功构建了人骨形成蛋白 -7(BMP -7)毕赤酵母表达重组子。结论 :为今后基因工程大量生产BMP -7打下基础。
OBJECTIVE: To construct a recombinant plasmid of Pichia pastoris from bone morphogenic protein-7 (BMP-7). METHODS: The coding sequence of human BMP-7 mature peptide was amplified by PCR using the recombinant plasmid pBV220 / BMP-7 stored in our laboratory as a template. The coding sequence of human BMP-7 mature peptide was cloned into pGEM-Teasy vector by A-T ligation and cloned into Pichia pastoris secretory expression vector pPIC9, the recombinant plasmid was linearized PEG into Pichia strain GS115, PCR screening of transformants. Results: DNA sequence analysis and PCR identification showed that recombinant human Porphyrin-7 (BMP-7) Pichia pastoris was successfully constructed. Conclusion: It laid the foundation for mass production of BMP-7 in future genetic engineering.