柑橘mRNA差异显示技术体系建立  

Establishing the System of Citrus mRNA Differential Display Technology

作　　者：刘凯[1,2] 曾继吾[1] 夏瑞[1] 陈金印[2] 易干军[1] 黄永红[1] 周碧容[1] 

机构地区：[1]广东省农业科学院果树研究所,广东广州510640 [2]江西农业大学农学院,江西南昌330045

出　　处：《基因组学与应用生物学》2009年第2期339-344,共6页Genomics and Applied Biology

基　　金：广东省科技计划项目(2008A020100022;2008A040102002);国家科技支撑计划课题(2008BAD92B05)资助

摘　　要：明柳甜橘是从春甜桔自然芽变中选育出来的新品种,本研究以春甜橘和明柳甜橘的叶片为试验材料,首先提取春甜橘和明柳甜橘叶片的总RNA,经3种锚定引物(HT11A,HT11C,HT11G)反转录成3类cDNA,再利用相应的锚定引物和34个随机引物(HAP1-HAP34)组成引物对,反转录cDNA进行差异显示PCR扩增,将扩增产物经6%的变性聚丙烯酰胺凝胶电泳后进行银染,获得了比较清晰的差异条带,然后回收差异条带并进行克隆,初步分离获得一些春甜橘和明柳甜橘差异基因,建立了柑桔mRNA差异显示技术,为进一步分析导致柑橘芽变的分子机理奠定基础。Mingliu tianju is a new variety was selected from Chuntianju nature mutant. In this study, the leaves of Chuntianju and Mingliu tianju are for test materials. First of all, total RNA is extracted from the leaves of Chuntianju and Mingliu tianju, using three kinds of anchor primers （HT11A, HT11C, HT11G） reverse transcription in-to three cDNA, then using the appropriate anchor primers and 34 random primers （HAPI-HAP34） composed of primers, the cDNA for reverse transcription is used for PCR amplification of differential display. The PCR product is confirmed by 6% denaturing polyacrylamide gel electrophoresis followed by silver staining, access to a relatively clear-cut difference between bands. And then recovering differential bands and cloning. Preliminary isolating and getting some of differential genes between Chuntianju and Mingliu tianju. At last, establishing the Citrus mRNA differential display, for further analysis led to Citrus mutant lay the foundation for the molecular mechanism.

关 键 词：柑橘 芽变 mRNA差异显示(DDRT-PCR) 

分 类 号：S666[农业科学—果树学]