微拟球藻硫脂酶基因克隆及其在大肠杆菌中的表达  

Cloning of a Thioesterase Gene from Nannochloropsis oceanica and Its Expression in Escherichia coli

作　　者：王丽丽[1] 朱葆华[1] 杨官品[2] 潘克厚[1] 

机构地区：[1]中国海洋大学海水养殖教育部重点实验室,山东青岛266003 [2]中国海洋大学海洋生命学院,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2015年第10期31-35,共5页Periodical of Ocean University of China

基　　金：国家自然科学基金项目(31372518);国家重点基础研究发展计划项目(2011CB200901);国家科技支撑计划项目(2011BAD14B01)资助

摘　　要：微拟球藻(Nannochloropsis oceanica)基因组序列注释的硫脂酶基因(NoTE)长1 314bp,其编码的硫脂酶长437氨基酸,与其他物种硫脂酶序列相似性较低。为验证其功能,将NoTE克隆到原核表达载体pET-30a上并在大肠杆菌BL21(DE3)中实现了表达。聚丙烯酰胺凝胶电泳(SDS-PAGE)检测到预期大小的蛋白质条带。气相色谱(GC)分析显示,表达NoTE未改变大肠杆菌脂肪酸种类,但改变了其总脂肪酸含量和脂肪酸比例,其C14∶1、C16∶0、C16∶1、C18∶1及总脂肪酸含量较不含重组质粒菌株分别提高了11%、18%、32%、49%和30%。研究结果证明了克隆的硫脂酶基因能在大肠杆菌中表达并具有相应的酶活性。The thioesterase gene annotated on Nannochloropsis oceanica genome sequence is 1 314 bp in length.The deduced Nannochloropsis thioesterase,437 aa in length,was phylogenetically far from those of other species in sequence.In order to verify the function of Nannochloropsis thioesterase gene（NoTE）,it was isolated from N.oceanica,inserted into pET-30 aand expressed in E.coli BL21（DE3）.SDS-PAGE detected a protein band with expected size.GC analysis revealed that expressing NoTEin E.coli BL21（DE3）didn′t change the components of fatty acids,but increased the contents of C14∶1,C16∶0,C16∶1and C18∶1and total fatty acids by 11%,18%,32%,49%and 30%,respectively.These findings indicated that the isolated gene expressed successfully in E.coli BL21（DE3）and functioned as a thioesterase gene.

关 键 词：微拟球藻 硫脂酶 脂肪酸 大肠杆菌 原核表达 

分 类 号：Q341[生物学—遗传学]