茶树生长素抑制蛋白基因CsARP1的克隆与表达分析  

CLONING AND EXPRESSION ANALYSIS OF AUXIN-REPRESSED PROTEIN GENE CsARP1 IN TEA PLANT(Camellia sinensis)

作　　者：王新超[1] 马春雷[1] 杨亚军[1] 姚明哲[1] 金基强[1] 

机构地区：[1]中国农业科学院茶叶研究所茶树资源和改良研究中心/国家茶树改良中心,浙江杭州310008

出　　处：《核农学报》2011年第5期910-915,921,共7页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金项目(30872059);现代农业(茶叶)产业技术体系项目(CARS-23)

摘　　要：从茶树休眠芽抑制消减杂交文库中分离得到生长素抑制蛋白基因的3'-片段,以休眠芽为材料,利用RACE技术克隆了其cDNA全长,并利用荧光定量PCR研究了该基因在不同休眠阶段芽的相对表达量。结果从茶树休眠芽中获得一个全长为711bp的生长素抑制蛋白基因CsARP1(GenBank登录号为HQ225758)。该基因开放阅读框为357bp,编码118个氨基酸,推测的蛋白质分子量为12.82KD,等电点约为9.57。多序列比对结果显示,该基因编码的氨基酸序列与其他植物的ARP蛋白序列相似性达到70%以上,具有生长素抑制基因家族的保守结构域。荧光定量PCR结果表明,CsARP1基因在休眠阶段表达量较高,而在解除休眠(萌发)后表达量较低,说明CsARP1基因可能与茶树芽休眠有关。A 3′-end gene fragment of auxin-repressed protein gene（ARP） was screened from the tea plant dormant bud suppression subtractive hybridization（SSH） library,its full-length cDNA sequence was cloned through rapid amplification of cDNA ends（RACE）,and its relative expression quantity in different stages of dormant buds was analyzed by real-time fluorescence quantitative PCR.The full length of the auxin-repressed protein gene,named CsARP1,was 711bp（GenBank accession No.HQ225758） and contained a 357bp open reading frame（ORF） encoding a 118 amino acid residues,and its 3′ untranslated region was an obvious polyadenylation signal.The deduced protein molecular weight was 12.82kD and its theoretical isoelectric point was 9.57.Sequence alignment of the deduced amino acids of CsARP1revealed a high degree of similarity with other members of plant ARP and had a typical domain characteristic.The results of real-time quantitative PCR showed that the CsARP1gene was expressed at a higher level in dormant buds than in sprouting buds.It suggests that the expression of CsARP1gene is correlated to the bud dormancy transition.

关 键 词：茶树 生长素抑制蛋白基因CsARP1 RACE 序列分析 实时荧光定量PCR 

分 类 号：S571.1[农业科学—茶叶生产加工]