水稻几丁质酶基因在大肠杆菌中的表达、纯化与活性分析

The Expression, Purification and Activity Analysis of the Rice Chitinase Gene in Escherichia coli

摘要:

将克隆并确定序列的水稻(Oryza sativa L．Cpslo17)几丁质酶基因Oschi的cDNA序列(此序列已在GenBank中注册, 登录号为EU045451)插入原核表达载体pGEX-4T-1中。经酶切、序列鉴定分析后, 用该重组质粒转化大肠杆菌BL21(DE3); 经IPTG诱导获得表达, 并对Oschi蛋白表达条件进行了优化; 在大肠杆菌中表达的几丁质酶经纯化后在一定的浓度、一定的反应时间内能高效地分解几丁质。

Abstract:

The cloned cDNA sequence of rice (Oryza sativa L．Cpslo17) chitinase gene Oschi was cloned, (which was registered in GenBank, the accession number: EU045451) ligated with the expression vector pGEX-4T-1, and transformed into E.coli BL21(DE3). The expression of Oschi was induced by IPTG, and the conditions were optimized. After purification the in vitro activity of Oschi chitinase was analyzed, and the results indicated that it could efficiently degrade chitin.