不同的引导肽对丝状噬菌体基因8蛋白展示外源多肽的影响

高等学校化学学报 ›› 2005, Vol. 26 ›› Issue (6): 1076.

不同的引导肽对丝状噬菌体基因8蛋白展示外源多肽的影响

王丽, 华攀玉, 高瑞娟, 杨琼, 宋金娜

东北师范大学生命科学学院遗传与细胞研究所, 长春130024

收稿日期:2004-06-21 出版日期:2005-06-10 发布日期:2005-06-10
通讯作者: 王　丽(1957年出生),女,博士,教授,从事生物化学及细胞生物学方面的研究.E-mail:wangli@nenu.edu.cn E-mail:wangli@nenu.edu.cn
基金资助:
国家自然科学基金(批准号:30471602);吉林省科技发展计划项目(批准号:970556-1);国家教育部留学回国人员科研项目资助.

Effects of Different Leader Peptides on the Foreign Peptide Displayed on Gene 8 Protein of Filamentous Phage

WANG Li, HUA Pan-Yu, GAO Rui-Juan, YANG Qiong, SONG Jin-Na

Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China

Received:2004-06-21 Online:2005-06-10 Published:2005-06-10

摘要/Abstract

摘要： 为了研究不同的引导肽对在基因8蛋白上展示外源多肽的影响,利用基因工程方法将一短肽插入基因8蛋白的N端,并将引导多肽去除或用基因3蛋白的引导肽序列替代基因8的引导肽序列.采用放射性脉冲追踪技术检测不同的引导肽对在基因8蛋白上展示外源多肽的作用.结果表明,无引导肽序列的引导,蛋白前体无法向成熟蛋白转化.基因3蛋白的引导肽可以引导展示有外源多肽的基因8蛋白,完成从前体向成熟蛋白的转化,但转化率明显下降.研究结果对阐明噬菌体外壳蛋白在大肠杆菌中的跨膜机制具有重要的意义.

关键词: 基因8蛋白, 引导肽序列, 噬菌体展示, 外源多肽

Abstract: Phage Display Technique was developed in 1985, it were applied in various biological fields. Phage can be exploited to display peptides, protein domains and even whole protein on its surface. Gene 3 protein and gene 8 protein are both the coat proteins of filamentous bacteriophage. Both of them were synthesized by using the precursors(pre-coats). Through different transmembrane pathways, they were inserted into the inner membrane of Escherichia coli cell and cleaved into mature proteins by the leader peptidase. To investigate the importance of the leader peptides, a small peptide was incorporated into the N-terminal region of gene 8 protein by using gene engineering methods. And for the first time, the leader peptide was deleted or replaced with the leader peptide of gene 3 protein. Pulse-chase analysis demonstrates the effects of different leader peptides on the polypeptide displayed on the N-terminal of gene 8 protein. The results showed that the precursor couldn't be converted into mature coat protein without the aid of leader peptide, while the leader peptide of gene 3 protein made gene 8 protein containing a foreign peptide become mature protein. These results have an important significance for the further studies of transmembrane mechanism of coat proteins of filamentous bacteriophage in E. coli.

Key words: Gene 8 protein, Leader peptide sequence, Phage display, Foreign peptide

中图分类号:

Q522⁺.4

TrendMD:

王丽, 华攀玉, 高瑞娟, 杨琼, 宋金娜. 不同的引导肽对丝状噬菌体基因8蛋白展示外源多肽的影响. 高等学校化学学报, 2005, 26(6): 1076.

WANG Li, HUA Pan-Yu, GAO Rui-Juan, YANG Qiong, SONG Jin-Na. Effects of Different Leader Peptides on the Foreign Peptide Displayed on Gene 8 Protein of Filamentous Phage. Chem. J. Chinese Universities, 2005, 26(6): 1076.

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