平邑甜茶非共生血红蛋白基因GLB1的克隆、表达及转化研究  

Study on Cloning,Expression and Transformation of Nonsymbiotic Hemoglobin Gene GLB1 from Malus hupehensis Rehd

作　　者：史兴征[1] 彭福田[1] 王新亮[1] 王兆燕[1] 赵玉[1] 

机构地区：[1]山东农业大学园艺科学与工程学院/作物生物学国家重点实验室,山东泰安271018

出　　处：《山东农业科学》2010年第6期5-9,共5页Shandong Agricultural Sciences

基　　金：山东省自然科学基金资助项目(Y2007D08);十一五科技支撑重点项目(2008BADA4B05)

摘　　要：根据GenBank上苹果GLB1基因的EST序列设计引物,利用PCR技术克隆到平邑甜茶GLB1同源基因MhGLB1(GenBank注册号:GQ423619),该基因包含一个477 bp的开放阅读框,编码158个氨基酸,分子量为17.8 ku。成功构建了35S∷MhGLB1正义表达载体,并对"S以12粉"番茄进行农杆菌介导的遗传转化。荧光定量PCR结果显示,MhGLB1在根、茎、叶中均有所表达,但在根中的表达量最高;NO3-和SNP处理能够诱导根中MhGLB1的表达。Based on the EST sequence of GLB1 from apple in GenBank, the primers were designed for PCR, and then the gene MhGLB1 was cloned from Malus hupehensis, whose accession number was GQ423619 in GenBank. The full - length cDNA contained a 477 bp open reading frame encoding a 17.8 ku protein with 158 amino acids. Its sense expression vector was also constructed as 35S ：： MhGLB1, which could be successfully used for genetic transformation mediated by Agrobacterium tumefaciens into tomato. The quantitative real - time PCR analysis showed that MhGLB1 expressed in roots, stems and leaves of Malus hupehensis, but the expression level was the highest in roots. The treatments of NO3- and SNP could induce its expression in roots.

关 键 词：平邑甜茶 MhGLB1 基因克隆 转基因 

分 类 号：Q78[生物学—分子生物学] S661.4[农业科学—果树学]