杉木木质化过程特异表达基因文库的构建与分析  

Construction and Analysis of Differentially Expressed cDNA Library from Differentiating Xylem of Chinese Fir

作　　者：王桂凤[1] 高燕[1] 杨立伟[1] 徐进[1] 席梦利[1] 施季森[1] 

机构地区：[1]林木遗传与基因工程江苏省和国家林业局重点实验室,南京林业大学,南京210037

出　　处：《分子植物育种》2006年第4期489-492,共4页Molecular Plant Breeding

基　　金：国家973项目（G199901600004）第四课题项目资金资助.

摘　　要：研究木质部木质化过程及其生物学特性对于了解木本植物水分和矿物质的吸收及运输机制、木材形成的遗传控制、改良材性及提高木材生长量具有十分重要的意义。为了获得杉木木质化过程中特异表达的基因,本研究利用抑制差减杂交技术,以杉木突变体独干杉无性系为测试方(Tester),正常的句容0号无性系为驱动方(Driver),构建了正向和反向两个差减文库,分别获得了618和409个克隆。利用通用引物T7和SP6进行PCR及EcoRⅠ酶切鉴定文库重组子,结果证明所构建的文库符合要求,具有代表性。该文库的成功构建为获得杉木木质化过程中特异表达的基因及深入了解木本植物木质化的机理奠定了基础。Wood formation （xylogenesis） is a critical developmental process for woody plants. The study ofxylogenesis and its biological characters is very important not only to understand the mechanism of transport and absorption of water and minerals but also to genetically control wood formation and improve wood properties and growth mass. To isolate the differentially expressed genes in xylogenesis of Chinese fir, a forward and a reverse subtractive cDNA libraries were constructed by using suppression subtractive hybridization method in this study, SSH was performed by using the cDNA from the mutant Duganshan （branchless） clone as Tester and the cDNA from the normal clone No.J0 as Driver, obtained 618 and 409 clones respectively. Recombinants were identified by using PCR with universal T7 and SP6 primers and using EcoR I digestion. The results showed that the subtractive cDNA libraries were successful and representative. The cDNA clones in the libraries have made a basis for the isolation of differentially expressed genes in xylogenesis and further understanding the mechanism of wood formation for woody land plants.

关 键 词：杉木 木质化 抑制差减杂交 特异性表达基因 

分 类 号：S791.27[农业科学—林木遗传育种]