利用抑制差减杂交技术分离棕色棉纤维特异表达基因  

Isolation of Specifically Expressed Genes in Brown Colored Cotton by Suppression Subtractive Hybridization

作　　者：李艳军[1] 张新宇[1] 杨会娜[1] 田新惠 孙杰[1] 

机构地区：[1]石河子大学农学院新疆兵团绿洲生态农业重点实验室,石河子832003

出　　处：《分子植物育种》2009年第1期194-198,共5页Molecular Plant Breeding

基　　金：国家973前期专项(2007CB116210);兵团科技局基础研究资金项目(2006JC02);石河子大学高层次人才启动资金项目(RCZX200517)资助

摘　　要：采用棕色棉新彩棉5号为Tester,其白色近等基因系NIL-C5为Driver,通过抑制差减杂交法(suppressive subtractive hybridization,SSH)构建了一个棕色棉纤维色素差异表达基因差减文库。从文库中随机挑选400个克隆,将这些克隆的PCR产物点膜进行反向Northern杂交检测,挑选20个差异明显的克隆进行序列分析,结果表明:有5个cDNA片段与已知的功能基因序列相似,其中1个cDNA片段与合成色素的重要酶基因4-香豆酸:CoA连接酶(4CL)基因有较高的同源性,其它cDNA片段在数据库中未找到同源序列。A subtracted eDNA library was constructed via suppressive subtractive hybridization （SSH）, in which the brown colored cotton Xincaimian 5 was utilized as tester while driver is its white colored susceptible near-isogenie line NIL-C5. Total 400 clones were identified from screening of the subtracted library by PCR, and further verified by Reverse Northern Dot-Blot. 20 differentially expressed clones were selected for sequencing analyses, of which the results were aligned with GenBank databases via BLASTX programs. It is shown that 5 of 20 candidates were identity with known genes in GenBank, 1 of them showed significant similarity to 4CL, a important gene for pigment synthesis, and no similar sequence were found in GenBank for the other fragments.

关 键 词：彩棉 抑制扣除杂交 差异基因 

分 类 号：S562[农业科学—作物学]