|
|||||
|
|
| Bacillus subtilis FS321的α-淀粉酶基因的克隆与表达 | |
| 引用本文： | 施碧红,温建新,吴伟斌,施巧琴,吴松刚.Bacillus subtilis FS321的α-淀粉酶基因的克隆与表达[J].中国农学通报,2011,27(33):194-198. |
| 作者姓名： | 施碧红 温建新 吴伟斌 施巧琴 吴松刚 |
| 作者单位： | 1. 福建师范大学教育部工业微生物工程研究中心,福州350108;福建师范大学生命科学学院,福州350108 2. 福建师范大学生命科学学院,福州,350108 3. 福建师范大学教育部工业微生物工程研究中心,福州,350108 |
| 基金项目： | 福建省自然科学基金项目(2009J01128); 福建省科技重点项目(2009N0033) |
| 摘 要： | 耐热α-淀粉酶被广泛用于食品等诸多行业，本研究从中国北方高温堆肥分离的枯草芽孢杆菌FS321中克隆了一中度耐热α-淀粉酶基因，并实现在大肠杆菌BL21（DE3）中的表达。通过PCR技术克隆Bacillus subtilis FS321的α-淀粉酶编码基因(BSA)，该基因全长1980 bp。并构建重组表达质粒pET-28a/BSA，转化大肠杆菌E. coli BL21(DE3)，经IPTG诱导表达，SDS-PAGE检测到大小约为73.0 kDa的重组融合蛋白，可溶性淀粉平板检测结果表明BSA在大肠杆菌中实现了有效表达。该重组α-淀粉酶的最适反应温度为50℃，最适反应pH为7.5。 |
| 关 键 词： | 气相色谱分析 气相色谱分析 |
| 收稿时间： | 2011/9/19 0:00:00 |
| 修稿时间： | 2011/10/21 0:00:00 |
Cloning and Expression of the α-amylase Gene from Bacillus subtilis FS321 |
|
| Shi Bihong , Wen Jianxin , Wu Weibin , Shi Qiaoqin , Wu Songgang.Cloning and Expression of the α-amylase Gene from Bacillus subtilis FS321[J].Chinese Agricultural Science Bulletin,2011,27(33):194-198. | |
| Authors: | Shi Bihong Wen Jianxin Wu Weibin Shi Qiaoqin Wu Songgang |
| Institution: | Shi Bihong1,2,Wen Jianxin2,Wu Weibin2,Shi Qiaoqin1,Wu Songgang1 (1Engineering Research Center of Industrial Microbiology,Ministry of Education,Fujian Normal University,Fuzhou 350108,2College of Life Sciences,Fuzhou 350108) |
| Abstract: | The thermostable α-amylase was widely used in the fields of food industry etc., in the present study, a moderate thermostable α-amylase gene was cloned from Bacillus subtilis FS321, which was isolated from the dunghill soil in the northern of China, and expressed in Escherichia coli BL21 （DE3）. The α-amylase gene from B.subtilis FS321 （BSA） was cloned using PCR method, the complete ORF of BSA was in a size of 1980 bp. The recombinant expression vector of pET-28a/BSA was constructed, and transformed to E. coli BL21 （DE3）. A fusion protein with a size of about 73.0 kD displayed in SDS-PAGE and the clear zones developed in the soluble starch plate demonstrated that the BSA was functional expressed in E. coli after induction by IPTG. The optimum reaction temperature of the recombinant α-amylase was of 50℃ , and optimum reaction pH of 7.5. |
| Keywords: | Bacillus subtilis α-amylase cloning expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |
| 点击此处可从《中国农学通报》浏览原始摘要信息 | |
| 点击此处可从《中国农学通报》下载免费的PDF全文 | |