β-半乳糖苷酶在枯草芽孢杆菌中的分泌表达

被引:9
作者
陈晓月 [1 ]
金宁一 [2 ]
邹伟 [2 ]
海洋 [2 ]
马海利 [1 ]
李昌 [1 ]
屈勇刚 [1 ]
机构
[1] 吉林大学畜牧兽医学院
[2] 中国人民解放军军事医学科学院军事兽医研究所
关键词
β-半乳糖苷酶; 枯草芽孢杆菌; 分泌表达;
D O I
10.13523/j.cb.20080519
中图分类号
Q78 [基因工程(遗传工程)];
学科分类号
071007 ; 0836 ; 090102 ;
摘要
为检测本课题组构建的枯草芽孢杆菌分泌表达载体pGPST中启动子和信号肽的活性,将β-半乳糖苷酶基因插入表达载体的多克隆位点,构建含有β-半乳糖苷酶基因的重组质粒pGPST-lacZ,重组质粒转化枯草芽孢杆菌,分别测定液体培养基中上清液和菌体沉淀中β-半乳糖苷酶的活性。培养上清液中的酶活在22h达到峰值,约为26mU/ml,之后开始下降;菌体沉淀中的酶活在14h最高,达6mU/ml,而阴性对照pGPST没有检测到酶活性。结果表明分泌型表达载体中的启动子和信号肽具有较好的活性,能实现异源基因在枯草芽孢杆菌中的分泌表达。本试验为该表达载体的进一步研究和应用提供重要的数据资料,也为外源基因在枯草芽孢杆菌中的表达提供参考资料。该载体将在研究外源基因在枯草芽孢杆菌中表达发挥重要作用。
引用
收藏
页码:111 / 115
页数:5
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