【摘要】 目的构建恶性疟原虫主要裂殖子表面蛋白1C末端相对分子质量1.9万片段(MSP1-19)毕氏酵母真核表达克隆。方法利用PCR技术扩增出MSP1-19,插入pPIC9载体中,鉴定正确的MSP1-19基因,插入毕氏酵母分泌型表达载体pPIC9k中,DNA测序鉴定序列的正确性,转化酵母细胞。结果筛选出的阳性克隆为MSP1-19表达克隆。结论用分子生物学方法重组构建的MSP1-19毕氏酵母真核表达克隆,为高效表达MSP1-19及其活性鉴定奠定了基础。更多还原

【Abstract】 Objective To construct MSP1-19 gene eukaryotic expression clones in pichia pastoris. Methods MSP1-19 genes were amplified by PCR technique from recombinant vector pPIC9/PFCP-2 and inserted into cloning vector pPIC9.Recombinant expression vectors were constructed by ligation with pPIC9k vectors.The validity of these sequences were confirmed by automatic DNA sequencing.By using electroporation transformation,recombinant expression vectors containing MSP1-19 genes were transformed into pichia GSll5 cells. Results The positive clones were identified with recombinant MSP1-19 gene clones. Conclusion The construction of MSP1-19 gene expression clone with molecular biology is the basis of MSP1-19 expression in pichia pastoris.更多还原