不结球白菜抗坏血酸过氧化物酶基因克隆及其原核表达载体构建  

Cloning and Construction of Prokaryotic Expression Vector for APX Gene from Non-heading Chinese Cabbage

作　　者：马成英[1] 侯喜林[1] 刘同坤[1] 李俊星[1] 张蜀宁[1] 李英[1] 

机构地区：[1]南京农业大学园艺学院,农业部南方蔬菜遗传改良重点开放实验室,江苏南京210095

出　　处：《江苏农业学报》2010年第4期790-795,共6页Jiangsu Journal of Agricultural Sciences

基　　金：国家“973”计划项目(2009CB119001-04)

摘　　要：参照拟南芥APX基因保守域序列设计引物,扩增出不结球白菜抗坏血酸过氧化物酶(APX)基因的核心片段,结合RACE技术获得该基因的5′端序列和3′端序列,利用DNAman软件经序列拼接获得1个全长为1 089bp的cDNA序列,该序列包括开放阅读框867 bp,编码288个氨基酸,蛋白质等电点5.52的相对分子量31 600。APX基因编码的氨基酸序列与拟南芥APX基因编码的氨基酸的同源性为81%。将APX基因片段连接到原核表达载体PGEX-4T-3,转化大肠杆菌BL21(DE3)后诱导重组蛋白质的表达,SDS-PAGE电泳结果表明,产生了预期大小的重组蛋白。Based on APX gene sequences of Aabidopsis thaliana in NCBI gene bank,an APX fragment of non-heading Chinese cabbage was amplified.The full length APX cDNA sequence was obtained by using RACE.The full length sequence of APX of non-heading Chinese cabbage showed 81% similarity with that of Arobidopsis and was named BC Apx1.BC Apx1 has an 867 bp coding region and encodes 288 amino acids.The putative protein of this gene has an isoelectric point of 5.52 and a calculated molecular weight of 31 600.BC Apx1 has been cloned into prokaryotic expression vector PGEX-4T-3 and transformed into the host BL21（DE3）.Results of SDS-PAGE showed that the specific fusion protein was successfully expressed by IPTG induction.

关 键 词：不结球白菜 抗坏血酸过氧化物酶基因 克隆 原核表达 

分 类 号：S634.3[农业科学—蔬菜学]