马铃薯腐烂茎线虫谷胱甘肽过氧化物酶基因(Dd-Gpx)的克隆与原核表达  

Cloning and Prokaryotic Expression of Dd-Gpx from Ditylenchus destructor

作　　者：刘洋[1] 何旭峰[1] 刘建喜[1] 丁中[1] 

机构地区：[1]湖南农业大学植物保护学院植物病虫害生物学与防控湖南重点实验室,长沙410128

出　　处：《生物技术通报》2015年第10期131-139,共9页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31071667)

摘　　要：旨在研究植物寄生线虫谷胱甘肽过氧化物酶(glutathione peroxidase,GPX)的结构和功能,通过RACE技术克隆了马铃薯腐烂茎线虫(Ditylenchus destructor)Gpx基因,并成功克隆到p ET-41b载体上进行融合表达。结果表明,该基因c NDA全长950 bp,含有1个681 bp的开放阅读框,共编码226个氨基酸。生物信息学分析显示,该蛋白N端有明显信号肽,属于分泌蛋白。将其克隆到p ET-41b载体在BL21(DE3)中进行原核表达,结果显示,在60 k D处有一条明显的带。经质谱鉴定,此条带有7个肽段与马铃薯腐烂茎线虫GPX匹配。表明重组Dd-GPX蛋白表达成功。To further understand the structure and function of glutathione peroxidase, a glutathione peroxidase gene was cloned from plant-parasitic nematode（Ditylenchus destructor）using RACE method, and the gene（Dd Gpx）was transferred into vector of p ET-41 b to have the fusion exp ression. The full-length c DNA of Dd Gpx was 950 bp containing an open reading frame of 681 bp and encoding 226 amino acids. Bioinformatics analysis indicated that the N terminal of the protein had obvious signal peptide, belonging to the secretory protein. The prokaryotic expression vector p ET-41b-Dd Gpx was constructed and then transformed into BL21（DE3）, there was a apparent band in 60 k D. Mass spectrum analysis indicated that there were 7 peptides matched with D. destructor GPX protein, implying that the recombinant protein of Dd-GPX was successfully expressed

关 键 词：马铃薯腐烂茎线虫 谷胱甘肽过氧化物酶 基因克隆 原核表达 

分 类 号：S432.45[农业科学—植物病理学]