| 血小板生成素在毕赤酵母中的表达 |
| |
| 引用本文： | 邢振兰,张红卫,郭树华,林泉,吴祖泽. 血小板生成素在毕赤酵母中的表达[J]. 山东大学学报(理学版), 2001, 36(1): 90-94 |
| |
| 作者姓名： | 邢振兰 张红卫 郭树华 林泉 吴祖泽 |
| |
| 作者单位： | 1. 山东大学生命科学院发育生物学研究所,
2. 中国军事医学科学院放射医学研究所, |
| |
| 摘 要： | 以人胎肝cDNA文库为模板，用PCR和DNA重组技术，将TPOcDNA克隆到pGEM
|
| 关 键 词： | 血小板生成素；毕赤酵母；基因重组；诱导表达 |
| 文章编号： | 0559-7234(2001)01-0090-05 |
| 修稿时间： | 2000-01-12 |
EXPRESSION OF HUMANTHROMBOPOIETIN IN PICHIA PASTORIS |
| |
| XING Zhen-lan,ZHANG Hong-wei,GUO Shu-hua,Lin Quan,WU Zu-ze. EXPRESSION OF HUMANTHROMBOPOIETIN IN PICHIA PASTORIS[J]. Journal of Shandong University, 2001, 36(1): 90-94 |
| |
| Authors: | XING Zhen-lan ZHANG Hong-wei GUO Shu-hua Lin Quan WU Zu-ze |
| |
| Affiliation: | XING Zhen-lan 1,ZHANG Hong-wei 1,GUO Shu-hua 2,LIN Quan 2,WU Zu-ze 2 |
| |
| Abstract: | By using the cDNA library from fetus liver and the technique of PCR and DNA recombinant ,TPO cDNAs was cloned into the pGEM -T vector and sequenced, then the target fragment was subcloned into a secretory expression vector pPIC9K to obtain the recombinant plasmid pPIC9K/IPO. We designed the TPO PCR primer to make the TPO cDNAs under the control of the methanol inducible alcohol oxidase promoter and make it be fused to the alpha mating factor (MFα) of Saccharomyces cerevisiae. The MFα can give signal for the secretion of protein. We got recombinant yeast KM71/pPIC9K/TPO by electrotransformation and secreened with G418. Then Pichia pastoris cells with pPIC9K/TPO were induced to express TPO. The molecular weight of approximately 65kD,as determined by SDS-PAGE and Western blot,is the exopected molecular weight foicorrectly processed and glycosylated protein. Scanning analysis indicated the expressed protein accounted up to 10.14％ of total secreted proteins. The result indicates that TPO can be successfully expressed in the methylotrophic yeast Pichia pastoris. |
| |
| Keywords: | thrombopoietin Pichia pastoris gene recombination inducible expression |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
