利用mRNA差异显示技术分离甜菜M_(14)品系特异表达基因的cDNA片段  

Extraction cDNA Fragments Specially Expressed in Lines M_(14) in Sugar Beet by mRNA Differential Display

作　　者：李海英[1] 马春泉[1] 于冰[1] 高传军[1] 张绍军[1] 张莹[1] 郭德栋[1] 

机构地区：[1]黑龙江大学生物技术实验室,哈尔滨150080

出　　处：《植物研究》2007年第4期465-468,共4页Bulletin of Botanical Research

基　　金：国家"863"高技术研究发展资助项目(863-101-04-01-07);国家转基因植物研究与产业化项目(JY03-A-2402);黑龙江省自然科学基金重点项目(ZJN-03-6);黑龙江大学博士后科研资金

摘　　要：二倍体栽培甜菜与白花甜菜杂交、进一步回交而获得的单体附加系M14,其染色体组成中除了含有18条栽培甜菜染色体外,还附加有一条野生白花甜菜第9号染色体,该附加染色体通过母本的传递率为96.5%;单体附加系传递率如此高的原因是因为M14中有无融合生殖基因的存在。本实验采用mRNA差异展示技术对甜菜无融合生殖品系M14和正常有性生殖的二倍体栽培甜菜A2Y花蕾减数分裂时期的基因表达进行了差异分析。采用GT15A,GT15G,GT15C3种锚定引物,共筛选了20个随机引物,通过RT-PCR检测,获得了6个阳性差异表达的cDNA片段,应用NCBI的BLASTx软件对测序结果进行同源序列、相似序列检索,为进一步克隆无融合生殖基因提供侯选cDNA片段。A monosomic addition line in sugar beet, which was designated as M14 was obtained by the wide cross between cultivated sugar beet （ Beta vulgaris L. ） and Beta corolliflora Zoss. M14 contained the 18 normal chromosomes of sugar beet plus the B. corolliflora chromosome 9. This alien chromosome in M14 had an average transmission frequency of 96. 5% through eggs, the reason it had so high transmission frequency was that it had apomixis genes. By mRNA differential display method, the study compared gene expression at the stage of meiosis in florescence between M14 and cultivated sugar beet （ Beta vulgaris L. ） A2Y which can carry out normal sexual reproduction. The anchored primer were GT15 A, GT15G, GT15C,and twenty arbitrary primers were used. By RT-PCR, six positive specific cDNA fragments were obtained. Analysis was performed on NCBI, and comparison the similarity with published sequences was carried out using the BLASTx program. These cDNA fragments were the candidate fragments to clone the apomixis genes.

关 键 词：无融合生殖 甜菜无融合生殖系M14 mRNA差异展示技术 

分 类 号：Q785[生物学—分子生物学]