长穗偃麦草EeHKT1;4基因的克隆与植物表达载体构建  

Gene cloning and construction of plant expression vector of EeHKT1;4 in Elytrigia elongata

作　　者：张琳[1] 郭强[1] 李杉杉[1] 毛培春[1] 田小霞[1] 孟林[1] ZHANG Lin GUO Qiang LI Shan-shan MAO Pei-chun TIAN Xiao-xia MENG Lin(Beijing Research and Development Center for Grass and Environment ,Beijing Academy of Agriculture and Forestry Science ,Beijing 100097,China)

机构地区：[1]北京市农林科学院,北京草业与环境研究发展中心,北京100097

出　　处：《草原与草坪》2017年第4期1-7,共7页Grassland and Turf

基　　金：国家自然科学基金项目(31272489);北京市农林科学院科技创新能力建设专项(KJCX20140103)资助

摘　　要：采用RT-PCR方法从长穗偃麦草(Elytrigia elongata)中克隆得到高亲和K+转运蛋白基因,命名为EeHKT1;4,全长cDNA序列为1 977bp,开放阅读框1 722bp,编码573个氨基酸,与一粒小麦(Triticum monococcum)TmHKT1;4-A2的氨基酸同源性为94%。系统进化树分析结果显示,EeHKT1;4与单子叶植物HKT1;4亲缘关系较近。利用In-Fusion技术,成功构建了pBI121-35SEeHKT1;4-Nos植物表达载体,为长穗偃麦草EeHKT1;4的耐盐功能鉴定奠定基础。A high-affinity K＋transporter gene was cloned fromElytrigia elongata using the RT-PCR technique.This Gene was designated as EeHKT1;4,and contained a 1 722 bp open reading frame and encoded 573 amino acids residues.The length of cDNA was 1 977 bp.The gene shared 94%of homology with TmHKT1;4-A2 from wheat（Triticum monococcum）.The gene has a closer genetic relationship with the HKT1;4from other monocotyledons using phylogenetic tree analyzing.In addition,the pBI121-35S-EeHKT1;4-Nos vector was constructed successfully using the In-Fusion technology.This would provide the important basis for further study in salt tolerance of EeHKT1;4 in Elytrigia elongata.

关 键 词：长穗偃麦草 高亲和K+转运蛋白(HKT) In-Fusion技术 载体构建 

分 类 号：S512[农业科学—作物学]