甜菜M14品系特异表达基因BvM14-MADSbox启动子的克隆及序列分析  

Cloning and analysis of the promoter of the gene BvM14-MADSbox specific expressed in Sugar Beet M14 Lines

作　　者：王玉婷[1] 曹洪祥[1] 马春泉[1] 王宇光[1] 郭锡伟[1] 李海英[1] 

机构地区：[1]黑龙江大学生命科学学院黑龙江省普通高等学校分子生物学重点实验室,哈尔滨150080

出　　处：《黑龙江大学自然科学学报》2009年第3期375-379,共5页Journal of Natural Science of Heilongjiang University

基　　金：国家自然科学基金资助项目(30871566);黑龙江省自然科学基金资助项目(C2007-37)

摘　　要：甜菜M14品系是带有白花甜菜染色体的栽培甜菜单体附加系,其染色体组成中除了包含18条栽培甜菜染色体外,还附加有白花甜菜第9号染色体,具有无融合生殖特性,无融合生殖能够固定杂种优势。在前期工作中,通过SSH、RACE等技术获得了甜菜M14品系特异表达基因BvM14-MADSbox的cDNA全长。通过6种限制性内切酶分别酶切M14品系基因组总DNA,再与相应接头连接,构建了6个DNA步移文库。根据基因BvM14-MADSboxcDNA序列设计引物,通过染色体步移技术克隆出该基因的上游DNA序列。经生物信息学分析,推测获得了BvM14-MADSbox基因上游1 718 bp的启动子序列,该序列含有启动子的基本元件TATA-box、CAAT-box,和ARE、ERE、HSE、ABRE、CGTCA-motif等重要的顺式调控元件及一些光反应相关元件。该结果为今后BvM14-MADSbox基因的时空特异性表达调控机制奠定基础。Sugar beet M14 with apomixis is a set of monosomic addition lines, constituted of the normal 18 Beta vulgaris L. chromosomes with the No. 9 chromosome of Beta corolliflora Zoss. Apomixis can be used to fix heterosis, which will be a novel approach in avoiding the tedious process of hybridization and seed production. In the earlier research, the cDNA full length of specific expression gene BvM14-MADSbox in M14 was obtained by the methods of SSH and RACE. In this experiment, genomic DNA from M14 was digested with six restriction enzymes respectively. A genomic walking cassette was ligated to the ends of the digested DNA fragments each, and then six DNA walking libraries were constructed. The 5′-flanking region of BvM14-MADSbox was amplified by LA-PCR from the libraries. 1 718bp sequence of promoter region was obtained using the bioinformatics analysis. The functional elements were also analysed. The BvM14-MADSbox gene promoter contains the basic elements： TATA-box,CAAT-box, some other important cis-regulatory elements, such as ARE,ERE,HSE,ABRE,CGTCA-motif and some relation to light elements are also found from the 5′-regulation region. It will be beneficial to the study of time-spatial specific expression of BvM14-MADSbox gene in the future.

关 键 词：甜菜M14品系 BvM14-MADSbox基因 启动子 序列分析 

分 类 号：S566.3[农业科学—作物学]