疟原虫_____](期刊论文)

恶性疟原虫MSP119毕氏酵母真核表达克隆的构建

http://www.100md.com 《青岛大学医学院院报》 2006年第1期

疟原虫,,],基因，MSP119；基因扩增；疟原虫，恶性；毕氏酵母；真核表达；聚合酶链反应,1材料与方法,2结果,3讨论,[参考文献]

     [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。

    [关键词] 基因，MSP119；基因扩增；疟原虫，恶性；毕氏酵母；真核表达；聚合酶链反应

    CONSTRUCTION OF MEROZOITE SURFACE PROTEIN1 CTERMINAL REGION(119) OF PLASMODIUM FALCIPARUM EUKARYOTIC EXPRESSION CLONEZHANG ZHONGGUANG, CHANG ZHISHANG, ZHAO HENGMEI, et al (Department of Parasitology, Qingdao University Medical College, Qingdao 266021, China)

    [ABSTRACT]ObjectiveTo construct MSP119 gene eukaryotic expression clones in pichia pastoris.MethodsMSP1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. ResultsThe positive clones were identified with recombinant MSP119 gene clones.ConclusionThe construction of MSP119 gene expression clone with molecular biology is the basis of MSP119 expression in pichia pastoris. ......

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