茶树功能候选基因电子克隆的可行性分析  

Feasibility Study of in Silico Cloning of Functional Candidate Genes from Tea(Camellia sinensis)

作　　者：孔祥瑞[1] 

机构地区：[1]福建省农业科学院茶叶研究所,福安355000

出　　处：《分子植物育种》2014年第2期332-337,共6页Molecular Plant Breeding

基　　金：国家科技支撑计划(2011BAD01B01);福建省农业科学院青年人才创新基金(2011QC-2);福建省农业科学院双百行动计划专项基金(sbmx1303-1)共同资助

摘　　要：为验证电子克隆技术获取茶树功能候选基因的可行性,本研究以可可的咖啡碱合成酶基因BCS1为探针,对构建的茶树EST数据库进行本地BLAST检索,获得与BCS1具有高度同源性的茶树EST序列26条。将26条茶树EST序列经程序CAP(contig assembly program)拼接,得到2条含有独立ORF的EST簇(Contig),分别命名为基因TCSnew1和TCSnew2。将这2个基因与GenBank中由实验方法克隆得到的茶树咖啡机合成酶基因TCS的cDNA进行核酸序列、推导氨基酸序列比对,及构建系统发育树进行3个基因推导氨基酸序列间的同源性分析。结果发现,茶树远缘物种的兴趣序列作为探针用于电子克隆获取茶树功能候选基因与实验克隆技术相同,是一条可行的技术途径。In order to verify the feasibility of the electronic cloning technology in tea functional gene cloning, theobroma cacao caffeine synthase gene 1 （BCS1） was chosen as a probe to blast the tea local EST database, as a result, 26 tea ESTs could been retrieved that were high homologous to BCS1, when these ESTs were assembled through CAP contig assembly program, two contigs had been obtained which have a perfect ORF respectively, and were named TCSnewl and TCSnew2. Then their nucleic acids and deduced amino acid sequences were compared with known gene TCS cDNA listed in the GenBank database cloned by experiment from tea, and the deduced amino acid sequences of above three genes were used to reconstructing a phylogenetic tree after accomplishing homologous analysis, the results showed that in silico cloning similar to experiment was an efficient way for cloning tea functional candidate genes by using distantly related species homologous gene as a probe.

关 键 词：茶树 电子克隆 咖啡碱合成酶基因 系统进化树 

分 类 号：S571.1[农业科学—茶叶生产加工]