摘要
采用同源克隆的方法获得金荞麦查尔酮合成酶基因(CHS)的保守片段554bp,进一步采用染色体步移法(genome-walking)和RT-PCR技术克隆到CHS基因的全长DNA序列和cDNA开放阅读框(ORF)序列。序列分析结果表明,金荞麦CHS基因DNA全长1650bp,含一个462bp的内含子;其cDNA编码区全长1188bp,编码395个氨基酸,命名为FdCHS,NCBI登录号为GU169470。该氨基酸序列与同为蓼科的掌叶大黄、虎杖CHS的氨基酸序列同源率分别达到94%和93%,且含有CHS活性位点和底物结合口袋位点等保守位点。
The conservative sequence of the chalcone synthase gene from Fagopyrum dibotrys 544 bp was isolated by homology cloning,and then the full-length DNA and cDNA ORF （open reading frame） sequences were cloned by genome-walking and RT-PCR,respectively.Sequence analysis showed that the CHS DNA of F.dibotrys was 1 650 bp,including one 462 bp intron.The coding region of cDNA was 1 188 bp,encoding 395 amino acids,designated as FdCHS and included the conserved sites of CHS such as active sites and substrate-binding pocket sites.The homology analysis of FdCHS with that of other plants showed that it was closely related to Rheum palmatum and Polygonum cuspidatum with 94% and 93% homology,respectively.The sequence accession number is GU169470 in GenBank.
出处
《草业学报》
CSCD
北大核心
2010年第3期162-169,共8页
Acta Prataculturae Sinica
基金
四川省科技厅科技攻关项目(04NG001-015
2006Z08-012)资助
关键词
金荞麦
查尔酮合成酶基因
克隆
序列分析
Fagopyrum dibotrys
chalcone synthase gene
cloning
sequence analysis
