Cloning, Characterization, and Molecular Application of a Beta-Agarase Gene from Vibrio sp. Strain V134
Cloning, Characterization, and Molecular Application of a Beta-Agarase Gene from Vibrio sp. Strain V134
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DOI：
10.1128/aem.02872-06
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发表时间：
2007-03
影响因子：
4.4
通讯作者：
Wei-Wei Zhang-Wei;Li Sun
中科院分区：
文献类型：
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作者：
Wei-Wei Zhang-Wei;Li Sun
Vibrio属海洋分离株V134可产生GH16家族β -琼脂酶。从V134中克隆出相应的琼脂酶基因agaV，并在大肠杆菌中有条件表达。酶活性分析表明，重组琼脂酶的最适温度和pH分别为40℃和7.0℃左右。实验证明AgaV具有两方面的用途：首先，作为一种琼脂糖水解酶，纯化后的重组AgaV可用于从琼脂糖凝胶中回收DNA；其次，作为一种分泌蛋白，AgaV在遗传水平上进行了探索，并作为一个报告基因用于构建分泌信号陷阱，该陷阱被证明是一种简单有效的分子工具，用于选择编码革兰氏阳性和革兰氏阴性细菌分泌蛋白的基因。
ABSTRACT V134, a marine isolate of the Vibrio genus, was found to produce a new beta-agarase of the GH16 family. The relevant agarase gene agaV was cloned from V134 and conditionally expressed in Escherichia coli. Enzyme activity analysis revealed that the optimum temperature and pH for the purified recombinant agarase were around 40°C and 7.0. AgaV was demonstrated to be useful in two aspects: first, as an agarolytic enzyme, the purified recombinant AgaV could be employed in the recovery of DNA from agarose gels; second, as a secretion protein, AgaV was explored at the genetic level and used as a reporter in the construction of a secretion signal trap which proved to be a simple and efficient molecular tool for the selection of genes encoding secretion proteins from both gram-positive and gram-negative bacteria.