苏云金芽孢杆菌几丁质酶基因chiA在枯草芽孢杆菌中的分泌表达Secretory Expression of Chitinase Gene chiA from Bacillus thuringiensis in Bacillus subtilis
宋光明;沈微;孙金凤;饶志明;方慧英;诸葛健;诸葛斌;
摘要(Abstract)：
[目的]研究来源于苏云金芽孢杆菌的几丁质酶基因chiA在枯草芽孢杆菌中的表达情况。[方法]以苏云金芽孢杆菌染色体DNA为模板,PCR扩增获取几丁质酶基因chiA,将其与枯草芽孢杆菌表达载体pHSG连接,构建重组菌,经IPTG诱导后,检测培养液中的几丁质酶活性。[结果]扩增得到几丁质酶基因chiA大小为2.5 kb。构建的重组菌对底物[4-MU-(GlcNAc)3]显示出一定的水解活性,培养液酶活约为2.8 U/ml,而pHSG空质粒转化子在同样条件下其培养液没有明显酶活。该重组酶的最适pH值为6.5,最适反应温度为50℃,与苏云金芽孢杆菌自身产生的几丁质酶性质一致。[结论]几丁质酶基因chiA能在枯草芽孢杆菌中成功表达,表达产物可成功分泌到细胞外。
关键词(KeyWords)： 苏云金芽孢杆菌;几丁质酶基因;枯草芽孢杆菌;分泌表达
基金项目(Foundation): 国家高技术研究发展计划(863计划,2006AA10Z307)资助项目
作者(Author): 宋光明;沈微;孙金凤;饶志明;方慧英;诸葛健;诸葛斌;
Email:
DOI: 10.13989/j.cnki.0517-6611.2008.26.057
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