金荞麦查尔酮合成酶基因CHS的克隆及序列分析

草业学报 ›› 2010, Vol. 19 ›› Issue (3): 162-169.

金荞麦查尔酮合成酶基因CHS的克隆及序列分析

蒙华,李成磊,吴琦^*,邵继荣,陈惠

四川农业大学生命科学与理学院,四川雅安 625014

收稿日期:2009-11-30 出版日期:2010-03-25 发布日期:2010-06-20
作者简介:蒙华(1985-),女,四川绵阳人,在读硕士。E-mail:lisa-mh@163.com
基金资助:
四川省科技厅科技攻关项目(04NG001-015,2006Z08-012)资助

Cloning and sequence analysis of the chalcone synthase gene (CHS) from Fagopyrum dibotrys

MENG Hua, LI Cheng-lei, WU Qi, SHAO Ji-rong, CHEN Hui

College of Life Science,Sichuan Agricultural University, Ya’an 625014, China

Received:2009-11-30 Online:2010-03-25 Published:2010-06-20

摘要/Abstract

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。

Abstract: 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.

中图分类号:

蒙华,李成磊,吴琦,邵继荣,陈惠. 金荞麦查尔酮合成酶基因CHS的克隆及序列分析[J]. 草业学报, 2010, 19(3): 162-169.

MENG Hua, LI Cheng-lei, WU Qi, SHAO Ji-rong, CHEN Hui. Cloning and sequence analysis of the chalcone synthase gene (CHS) from Fagopyrum dibotrys[J]. Acta Prataculturae Sinica, 2010, 19(3): 162-169.

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