北美海蓬子甜菜碱醛脱氢酶基因的克隆及表达载体构建  

Cloning of Gene from Salicornia bigelovii and its Expression Vectors Construction

作　　者：刘增美[1] 朱天艺[1] 龚一富[1] 王何瑜[1] 章丽[1] 

机构地区：[1]宁波大学海洋学院,浙江宁波315211

出　　处：《宁波大学学报（理工版）》2015年第4期16-19,共4页Journal of Ningbo University：Natural Science and Engineering Edition

基　　金：浙江省自然科学基金(LY13C020004);浙江省科技创新团队项目(2012R10029-07);宁波市科技攻关项目(2013C10018)

摘　　要：以北美海蓬子种子为材料,采用RACE技术获得北美海蓬子BADH基因的c DNA全长序列.结果表明:BADH基因c DNA全长为1 836 bp,其中开放阅读框(ORF)为1 503 bp,编码500个氨基酸,预测蛋白的分子量为54.5 k Da,理论等电点为5.31.推测出BADH氨基酸中含有甜菜碱醛脱氢酶家族高度保守的十肽序列(VTLELGGKSP)及与酶功能相关的半胱氨酸残基(Cys).序列比对和系统进化树显示,北美海蓬子与盐节木和盐穗木的亲缘关系最近,同源性达94%.并构建了植物表达载体p CAMBIA3301-BADH,将其成功导入到农杆菌EHA105中,为进一步研究该基因的功能奠定了基础.A full length cDNA sequence of BADH gene was cloned by RACE from Salicornia bigelovii seeds. The results show that the full length cDNA of BADH gene is 1 836 bp, which contains 1 503 bp ORF and encoded 500 amino acid with a putative molecular mass of 54.5 kDa and an isoionic point of 5.31 .The deduced amino acid sequence contains the conserved decapeptide sequence （VTLELGGKSP） and cysteine residue in betaine aldehyde dehydrogenase. Sequence comparison and phylogenetic tree analysis reveal that Salicornia bigeloviiwas is close to Halocnemum strobilaeeum and Halostachys easpica with 94% amino acids similarity. The plant expression vector pCAMBIA3301-BADH is successfully constructed, and the constructed vector is then transformed into Agrobacterium EHA105. The research is expected to lay a foundation for the further study on function ofBADH gene.

关 键 词：北美海蓬子 甜菜碱醛脱氢酶 基因克隆 载体构建 

分 类 号：Q781[生物学—分子生物学]