四川农业大学学报

• 研究论文 • 上一篇    下一篇

短小芽孢杆菌胶原蛋白酶基因cola的克隆及序列分析

赵海霞   

  1. 四川农业大学
  • 收稿日期:2011-08-09 修回日期:2011-11-09 出版日期:2011-12-25 发布日期:2011-12-26
  • 通讯作者: 赵海霞

Cloning and Sequence Analysis of Collagenase Gene Cola from Bacillus pumilus col-J

zhao haixia   

  1. sichuan agricultural university
  • Received:2011-08-09 Revised:2011-11-09 Online:2011-12-25 Published:2011-12-26
  • Contact: zhao haixia

摘要:

根据芽孢杆菌(Bacillus)胶原蛋白酶基因序列保守区域设计一对兼并引物,采用PCR法获得胶原蛋白酶产生菌短小芽孢杆菌(B. pumilus)col-J株胶原蛋白酶基因保守片段。序列分析表明,该片段与枯草芽孢杆菌(B.subtilis)基因组序列AL009126中编码胶原蛋白酶基因序列高度同源。参考该序列,获得了短小芽孢杆菌col-J株胶原蛋白酶基因全长开放阅读框序列。序列分析及生物信息学分析表明,该基因长930bp,推测为编码含309个氨基酸残基的胶原蛋白酶基因cola,理论分子量47.63KD,为非分泌性蛋白,仅有1个潜在的N-糖基化位点,密码子使用具有明显的偏爱性。

关键词: 短小芽孢杆菌, 胶原蛋白酶, 基因克隆, 序列分析

Abstract:

According to the conserved regions of Bacillus collagenase gene, a pair of degenerate primers were designed to amplify collagenase gene fragment from B. pumilus strain col-J with collagenolytic activity. Sequence analysis showed that the fragment sequence was highly homologous to a sequence encoding collagenase in the genome of B. subtilis AL009126 in GenBank.Based on the sequence, the collagenase gene full-length open reading frame cola was obtained from B. pumilus col-J strain. Sequence analysis and bioinformatics analysis showed that it was 930bp in length, presumably encoded a collagenase with 309 amino acid residues with 47.63KD theoretically. Furthermore, its encoding protein was a non-secreted protein with only one potential N-glycosylation site, and showed a strong usage bias.

Key words: Bacillus pumilus, Collagenase, Gene cloning, Sequence analysis