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| TIMP-2在毕赤酵母中的克隆与表达 | |
| 引用本文： | 闫训友,赵洪亮,张惟广,薛冲,刘志敏.TIMP-2在毕赤酵母中的克隆与表达[J].微生物学通报,2006,33(1):1-6. |
| 作者姓名： | 闫训友 赵洪亮 张惟广 薛冲 刘志敏 |
| 作者单位： | 1. 军事医学科学院生物工程研究所,北京,100071;廊坊师范学院生物系,廊坊,065000;西南农业大学食品科学学院,重庆,400716 2. 军事医学科学院生物工程研究所,北京,100071 3. 廊坊师范学院生物系,廊坊,065000 |
| 基金项目： | 国家重大科技专项平台课题基金资助（No.2002AA2Z345B） |
| 摘 要： | 为了克隆人基质金属蛋白酶组织抑制剂-2（TIMP-2）基因，并在Pichia pastoris中表达，根据GenBank上的TIMP-2的氨基酸序列和毕赤酵母偏爱密码子，通过化学合成和PCR相结合的方法获得了人的TIMP-2基因全长序列，构建了pPIC9-T2表达载体，电击转化到毕赤酵母，通过表型筛选和诱导表达得到蛋白表达工程菌，并对表达产物进行了分离纯化和生物学活性分析。 |
| 关 键 词： | TIMP-2 毕赤酵母 表达 活性 |
| 文章编号： | 0253-2654（2006）01-0001-06 |
| 收稿时间： | 2005-03-10 |
| 修稿时间： | 2005-05-26 |
Expression of Tissue Inhibitor of Metalloproteinase-2 (TIMP-2) in Pichia pastoris |
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| YAN Xun-You,ZHAO Hong-Liang,ZHANG Wei-Guang,XUE Chong,LIU Zhi-Min.Expression of Tissue Inhibitor of Metalloproteinase-2 (TIMP-2) in Pichia pastoris[J].Microbiology,2006,33(1):1-6. | |
| Authors: | YAN Xun-You ZHAO Hong-Liang ZHANG Wei-Guang XUE Chong LIU Zhi-Min |
| Institution: | 1.Insitute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071 ;2. Biology Department Lanfimg Normal College, Langfang 065000 ; 3. College of Food Science, Southwest Agricultural University Chongqing 400716 |
| Abstract: | To construct human tissue inhibitor of metallopmteiaase-2 and express it in P. pastoris. The sequence was designed according to the amino acid sequence of the TIMP-2 and high-level expression sequences of P. pastoris. We gained the TIMP-2 gene by artificially synthesis method. The gene was cloned into pPIC9 to construct a recombinant vector pPIC9-T2. The vector was transformed into P. pastoris by electroporation, through phenotype selection and induced with methanol we get the expression cell. Separation, purification and bioactivity analysis of tbe expressed products was performed. |
| Keywords: | TIMP-2 Pichia pastoris Express Activity |
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