| 恶性疟原虫MSP1-19毕氏酵母真核表达克隆的构建 |
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| 引用本文： | 张忠广,常志尚,赵恒梅,宫玉香.恶性疟原虫MSP1-19毕氏酵母真核表达克隆的构建[J].青岛大学医学院学报,2006,42(1):50-52. |
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| 作者姓名： | 张忠广 常志尚 赵恒梅 宫玉香 |
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| 作者单位： | 青岛大学医学院人体寄生虫学教研室,山东,青岛,266021 |
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| 基金项目： | 山东省青岛市科技局资助项目 |
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| 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。
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| 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 |
| 文章编号： | 1672-4488（2006）01-0050-03 |
| 收稿时间： | 2005-08-18 |
| 修稿时间： | 2005-12-08 |
CONSTRUCTION OF MEROZOITE SURFACE PROTEIN-1 C-TERMINAL REGION(I-19) OF PLASMODIUM FALCIPARUM EU-KARYOTIC EXPRESSION CLONE |
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| ZHANG ZHONG-GUANG, CHANG ZHI-SHANG, ZHAO HENG-MEI , et al.CONSTRUCTION OF MEROZOITE SURFACE PROTEIN-1 C-TERMINAL REGION(I-19) OF PLASMODIUM FALCIPARUM EU-KARYOTIC EXPRESSION CLONE[J].Acta Academiae Medicinae Qingdao Universitatis,2006,42(1):50-52. |
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| Authors: | ZHANG ZHONG-GUANG CHANG ZHI-SHANG ZHAO HENG-MEI |
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| Abstract: | Objective To construct MSP1-19 gene eukaryotic expression clones in pichia pastoris. Methods MSP1-19 genes were amplofied by PCR technique from recombinant vector pPICg/PFCP 2 and inserted into cloning vector pPICg. Recombinant expression vectors were constructed by ligation with pPICgk vectors. The validity of these sequencea were confirmed by automatic DNA sequencing. By using electroporation transformation, recombinant expression vectors containing MSP1-19 genes were transformed into piehia GSII5 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. |
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| Keywords: | genes MSP1-19 gene amplification plasmodium falciparum eukaryotic expression pichia pastoristpolymerase chain reaction |
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