日本落叶松咖啡酸-O-甲基转移酶基因LkCOMT的克隆及单核苷酸多态性分析  

Isolation and Single Nucleotide Polymorphisms Analysis of Caffeic Acid O-methyltransferase （LkCOMT） in Lar/x kaempferi

作　　者：易敏[1] 张守攻[1] 谢允慧[1] 孙晓梅[1] 

机构地区：[1]林木遗传育种国家重点实验室,中国林业科学研究院林业研究所,北京100091

出　　处：《林业科学研究》2013年第F10期52-59,共8页Forest Research

基　　金：国家“973”计划项目（2009CB119100）;国家863项目（2011AA100203）

摘　　要：依据日本落叶松转录组数据库检测到的咖啡酸-O-甲基转移酶（COMT）基因ESTs序列设计引物，分离得到一个编码该酶的新基因。其cDNA克隆全长为1350bp，包含长度为1092bp的开放阅读框，可编码364个氨基酸残基。序列分析显示，所推导的蛋白质氨基酸序列包含COMT基因5个所特有的保守元件，与海岸松PpCOMT的蛋白质氨基酸序列同源性达94％。在此基础上，利用DnaSP5．0软件对日本落叶松加株基因型个体的LkCOMT序列进行了单核苷酸多样性SNPs分析，共检测到92个SNP位点，SNP发生频率为1／17bp，多样性指数1T，为0．00780。在这些SNPs中，65个属于转换，27个属于颠换。在外显子区域，共检测到58个SNP位点，其中37个为错义突变，21个为同义突变。研究结果为Et本落叶松基因标记辅助育种提供了理论基础。The ESTs sequences of caffeic acid O-methyhransferase from Larix kaempferi transcriptome database, an cDNA clone encoding COMT, was isolated from L. kaempferi by gene-specific PCR amplification. The COMT clone was 1 350 bp in length with an open reading frame （ ORF, 1 092 bp） which would be capable to encode a protein of 364 AA. The sequence indicated that the deduced amino acid contained five highly conserved regions of COMT family, and shared 94% identity with PpCOMT from Pinus pinaster. The genomic sequences of LkCOMT in 40 indi- viduals were sequenced, and single nucleotide polymorphisms （SNPs） diversity of LkCOMT was analyzed using the software DnaSP5.0. A total of 92 SNPs were detected, and the frequency and diversity of SNPs （~rT） were 1/17 bp and 0. 007 80, respectively. There were 65 transition and 27 transversion of mutation types. A total of 58 SNPs were detected in the coding regions of LkCOMT, of which 15 were synonymous mutation and 37 were missense mutation.

关 键 词：日本落叶松 咖啡酸-O-甲基转移酶基因 基因克隆 单核苷酸多态性 

分 类 号：S791.223[农业科学—林木遗传育种]