河北大学学报(自然科学版) >

2018 , Vol. 38 >Issue 4: 403 - 409

DOI: https://doi.org/10.3969/j.issn.1000-1565.2018.04.010

纤维素降解菌N2-10菌株β-1,4-内切葡聚糖酶基因的克隆及表达

  • 狄聪颖 ,
  • 郭晓军 ,
  • 刘宏丽 ,
  • 郭威 ,
  • 朱宝成
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狄聪颖(1989—),女,河北河间人,河北农业大学在读硕士研究生. E-mail:dicy123@163.com

收稿日期: 2017-03-28

  网络出版日期: 2018-07-25

基金资助

河北省重点研发计划农业关键共性技术攻关专项(16226604D);河北省技术创新引导计划科技型中小企业技术创新资金专项(15C1303121015);沧州市科技支撑计划项目(161201007D);保定市科学技术研究与发展计划项目(16ZN007)

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Cloning and expression of β-1,4-endoglucanase gene of cellulose derading bacteria N2-10

  • DI Congying ,
  • GUO Xiaojun ,
  • LIU Hongli ,
  • GUO Wei ,
  • ZHU Baocheng
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  • 1.College of Life Science, Agriculture University of Hebei, Baoding 071000, China; 2.Hebei Zhongbang Biotechnology Limited Company, Baoding 071000, China

Received date: 2017-03-28

  Online published: 2018-07-25

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摘要

为探究枯草芽孢杆菌(Bacillus subtilis)N2-10菌株降解纤维素的机理,通过同源性比对设计兼并引物扩增菌株β-1,4-内切酶葡聚糖基因,并进行生物信息学分析,然后研究其在大肠杆菌中的表达情况.结果显示,该内切葡聚糖酶基因大小为1500 bp,共编码499个氨基酸,具有典型的纤维素酶结构,SDS-PAGE电泳检测其表观分子质量约为55 ku;刚果红染色显示,表达产物具有纤维素酶活性.

关键词： 纤维素降解菌; 枯草芽孢杆菌N2-10; 内切葡聚糖酶; 克隆; 表达

本文引用格式

狄聪颖 , 郭晓军 , 刘宏丽 , 郭威 , 朱宝成 . 纤维素降解菌N2-10菌株β-1,4-内切葡聚糖酶基因的克隆及表达[J]. 河北大学学报(自然科学版), 2018 , 38(4) : 403 -409 . DOI: 10.3969/j.issn.1000-1565.2018.04.010

Abstract

To validate the mechanism of cellulose degradation by Bacillus subtilis N2-10, through homology comparison,the degenerate primers was designed and the β-1,4-endoglucanase gene of B. subtilis N2-10 was amplified. By useing bioinformatics tools in analyzing the sequences of genes and proteins and considering that the expression of the β-1,4-endoglucanase gene in E.coli, the result shows that, this gene is 1 500 bp, which could totally encodle 499 amino acids. This study also found that it had the typical structure of cellulose. SDS PAGE analysis found that the apparent molecular weight of the expression product was about 55 ku. Congo red staining analysis found that the expression product had the activity of cellulose.

Key words： cellulose derading bacteria; Bacillus subtilis N2-10; endoglucanase; clone; expression

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