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参与紫云英共生固氮的1个上调表达结瘤素基因AsB6的分离与鉴定
引用本文:吴 梅,丑敏霞,李一星,陈大松,李友国.参与紫云英共生固氮的1个上调表达结瘤素基因AsB6的分离与鉴定[J].华中农业大学学报,2010,29(2):169-174.
作者姓名:吴 梅  丑敏霞  李一星  陈大松  李友国
作者单位:华中农业大学农业微生物学国家重点实验室,武汉,430070
基金项目:国家“973”计划课题(2010CB126502);;华中农业大学人才启动基金(2005XRC026)资助
摘    要:基于接种华癸中慢生根瘤菌(Mesorhizobiumhuakuii)7653R的紫云英(Astragalus sinicusL.)感染根与未接种对照根在转录水平上的差异,利用抑制差减杂交技术(suppression subtractive hybridization,SSH),建立了紫云英结瘤固氮过程中的差异表达cDNA文库,共获得527个有效克隆,其中增强或特异性表达的上调文库中包含341个克隆,抑制表达的下调文库包含186个克隆。对其中的1个上调表达cDNA克隆AsB6利用RACE(rapid amplification of cDNAends)方法获得了其基因全长序列,利用BLAST在线软件和GenBank,Gen-Bank EST和Tigr Porcine EST等数据库进行同源序列比较,表明AsB6与苜蓿的β-酮酯酰合成酶及依赖于SAM(S-adenosgl methiomine,S-腺苷蛋氨酸)的羧甲基转移酶具有66%的同源性,同时利用实时荧光定量PCR研究了该基因的时空表达特征。结果显示,接种根瘤菌7653R后,在紫云英感染根和根瘤中该基因的表达量显著增强,且在接种后根瘤形成早期和固氮中晚期表达...

关 键 词:抑制差减杂交  紫云英  上调基因  β-酮酯酰合成酶  共生固氮  
收稿时间:3/2/2009 12:00:00 AM
修稿时间:2009/11/12 0:00:00

Isolation and Identification of an Up-Regulated Nodulin Gene AsB6 Involved in Symbiotic Nitrogen Fixation of Astragalus sinicus L.
WU Mei,CHOU Min-xi,LI Yi-xing,CHEN Da-song and LI You-guo.Isolation and Identification of an Up-Regulated Nodulin Gene AsB6 Involved in Symbiotic Nitrogen Fixation of Astragalus sinicus L.[J].Journal of Huazhong Agricultural University,2010,29(2):169-174.
Authors:WU Mei  CHOU Min-xi  LI Yi-xing  CHEN Da-song and LI You-guo
Institution:State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China;State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China;State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China;State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China;State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China
Abstract:A cDNA library of Astragalus sinicus L.genes specifically expressed in infected roots by Mesorhizobium huakuii 7653R was generated by using a PCR-based suppressive subtractive hybridization (SSH) technique with two mRNA populations of infected and uninfected roots.A total number of approximately 527 SSH cDNA clones were obtained,including 341 up-regulated gene clones and 186 down-regulated gene clones.The full-length cDNA sequence of up-regulated AsB6 gene was obtained by RACE and analyzed through Blasting,...
Keywords:suppression subtractive hybridization  Astragalus sinicus  up-regulated gene  beta-ketoacyl synthase  symbiotic nitrogen fixation  
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