川草2号老芒麦(Elymus sibiricusL.)UV-B辐射敏感基因rbcL的克隆及其调控表达研究  

Isolation of the Genes Related to UV-B Radiation Sensitivity Using DDRT-PCR in Elymus sibiricus L

作　　者：何文兴[1] 李洪梅[2] 徐莺[1] 肖猛[1] 李群[1] 颜钫[1] 王胜华[1] 陈放[1] 

机构地区：[1]四川大学生命科学学院,成都610064 [2]济南大学化学化工学院,济南250022

出　　处：《中国生物工程杂志》2006年第12期56-62,共7页China Biotechnology

基　　金：国家"十五"科技攻关计划资助项目(2001BA606A-11)

摘　　要：rbcL是编码光合关键酶1,5-二磷酸核酮糖羧化酶(Rubisco)大亚基的基因。运用mRNA差异显示技术(DDRT-PCR)并通过5′RACE(rapidamplificationofcDNAends)从高原植物川草2号老芒麦(ElymussibiricusL.cv.‘chuancaoNo.2’)中获得了受增强UV-B辐射抑制的rbcL基因,该基因全长cDNA为1.51kb,开放阅读框(ORF)长1.434kb,编码477个氨基酸。氨基酸序列与Elymustrachycaulus中的Rubisco大亚基具有97%的同源性、与Triticumaestivum和Hordeumcomosum的Rubisco大亚基同源性均为98%。Northern杂交分析表明,增强UV-B辐射后6h,rbcL基因表达受到强烈抑制,处理后60h,其表达几乎完全被抑制,表明即使是长期生长在高原地区、强UV-B辐射条件下的高原物种,在受到较强的UV-B辐射后,其rbcL基因的转录也会受到抑制。rbcL is the gene which codes the large subunit of the photosynthetic enzyme, Rubisco. Its EST sequence, whose expression is inhibited by the enhanced UV-B radiation, was obtained from the leaf cDNA of alpine plant Elymus sibiricus L. cv. ‘chuancao No. 2＇ by reverse transcription-polymerase chain reaction （ RT- PCR） differential display method and a 1.S1 kb full-length cDNA was cloned through 5′RACE. The rbcL has a 1. 434 kb open reading frame （ORF） which encodes a precursor protein of 477 amino acid residues. The amino acid sequence exhibits 97%, 98% and 98% positional identity with its correspondent from Elymus trachycaulus Triticum aestivum,Hordeum comosum, respectively. Northern hybridization analysis on the rbcL expression during the time of enhanced UV-B radiation, showed that the mRNA level of the rbcL descended obviously after being exposed to enhanced UV-B radiation for 6h, and 60h later, the mRNA even couldn＇t be detected.‘ chuancao No. 2’ growing in the plateau area for a long time and being radiated strongly by the UV-B, the rbcL transcription of Elymus sibiricus L might be restrained like the plain plants, which are sensitive to UV-B.

关 键 词：RBCL基因 mRNA差异显示(DDRT-PCR) UV-B辐射 川草2号老芒麦 

分 类 号：S543[农业科学—作物学]