| 松材线虫纤维素酶基因的克隆及在大肠杆菌中的表达 |
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| 引用本文： | 任慧杰,郭道森,赵博光,李荣贵.松材线虫纤维素酶基因的克隆及在大肠杆菌中的表达[J].青岛大学学报(工程技术版),2011,26(1):59-65. |
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| 作者姓名： | 任慧杰 郭道森 赵博光 李荣贵 |
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| 作者单位： | 1. 青岛大学生物系,山东青岛,266071
2. 南京林业大学森林资源与环境学院,南京,210037 |
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| 基金项目： | 国家自然科学基金资助项目，山东省自然科学基金资助项目 |
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| 摘 要： | 采用PCR技术克隆了松材线虫纤维素酶的编码基因BXC46,克隆基因与已报道的纤维素酶基因BXC10（GenBank登录号：FJ598020.1）的同源性高达99%,将其克隆到表达载体pET-15b上构建了表达载体pET-15b-BXC。将该重组质粒转化大肠杆菌BL21（DE3）菌株构建工程菌,SDS-PAGE分析表明,IPTG诱导了纤维素酶在工程菌中表达。采用Ni2＋-NTA树脂亲和层析部分纯化了重组松材线虫纤维素酶,DNS法进行的活性测定表明,部分纯化的重组蛋白具有纤维素酶的活性,其最适温度为45℃,最适pH为4.0。
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| 关 键 词： | 松材线虫 纤维素酶 基因克隆 表达 |
Gene Cloning of Cellulase from Bursaphelenchus Xylophilus and Its Expression in Escherichia Coli |
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| REN Hui-jie,GUO Dao-sen,ZHAO Bo-guang,LI Rong-gui.Gene Cloning of Cellulase from Bursaphelenchus Xylophilus and Its Expression in Escherichia Coli[J].Journal of Qingdao University(Engineering & Technology Edition),2011,26(1):59-65. |
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| Authors: | REN Hui-jie GUO Dao-sen ZHAO Bo-guang LI Rong-gui |
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| Affiliation: | 1(1.Department of Biology,Qingdao University,Qingdao 266071,China;2.College of Forest Resources and Environment,Nanjing Forestry University,Nanjing 210037,China) |
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| Abstract: | A gene encoding cellulase BXC46 from Bursaphelenchus xylophilus was cloned by PCR,and showed 99% homology to the reported gene of cellulase BXC10(GenBank accession number FJ598020.1).The gene was cloned into pET-15b to construct expressing vector pET-15b-BXC.Then,this recombinant plasmid was introduced into E.coli BL21(DE3) to construct engineering bacterium.Recombinant cellulase was successfully expressed in engineering bacterium induced by IPTG as analyzed by SDS-PAGE.Recombinant protein was partially purified by Ni2+-NTA resin and showed good activity as determined by DNS method.The optimal temperature and pH of the recombinant cellulase were 45 ℃ and 4.0,respectively. |
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| Keywords: | Bursaphelenchus xylophilus cellulase gene cloning expression |
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