草莓半胱氨酸蛋白酶基因(FaCP)的克隆及表达分析  

Cloning and Expression Analysis of Cysteine Protease Gene(FaCP) in Strawberry(Fragaria × ananassa)

作　　者：朱海生[1,2,3] 陈敏氡[4] 温庆放[1,2,3] 林珲[1,2,3] 

机构地区：[1]福建省农业科学院作物研究所,福州350013 [2]福建省农业科学院蔬菜研究中心,福州350013 [3]福建省蔬菜工程技术研究中心,福州350013 [4]福建农林大学园艺学院,福州350002

出　　处：《农业生物技术学报》2013年第2期158-164,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.31101534);福建省自然科学基金项目(No.2010J0111);福建省农业科学院科技创新团队重点科研项目(No.CXTD2011-20)

摘　　要：半胱氨酸蛋白酶作为一种重要的水解蛋白酶,参与植物的许多生理过程。采用RT-PCR和RACE技术从草莓(Fragaria×ananassa)中克隆到半胱氨酸蛋白酶基因FaCP(GenBank登录号:JN979371),该基因cDNA全长1338bp,包含一个1062bp完整的开放阅读框(ORF),编码354氨基酸。生物信息学序列分析表明,FaCP开放阅读框编码的氨基酸序列与其他植物的CP蛋白同源性较高。Real-timePCR分析发现,FaCP基因在草莓果实、叶、根、茎和花萼中都有表达;在果实成熟过程中FaCP表达量逐渐增加,在粉红果最高,红果中略有下降。在草莓叶片中,随着叶片衰老,FaCP基因表达量增加明显。研究结果表明,FaCP可能参与了调控草莓果实的成熟及叶片衰老。Cysteine protease （CP） is one of the important hydrolysis protease, which widely participates in a variety of physiological processes of plants. The FaCP gene cDNA was cloned from strawberry （Fragaria x ananassa） using RT-PCR and RACE techniques. The cDNA sequence was 1 338 bp （GenBank accession number： JN979371）, including 1 065 bp of open reading frame （ORF）, which encoded a protein of 354 amino acids with a predicted molecular weight of 39 130 D and a hypothetical pI （isoelectric point） of 4.93. Homology analysis showed that the deduced CP protein was highly homologous to other CP proteins from different plant species. Phylogenetic analysis indicated that FaCP was more related to castor, trichocarpa, kiwi and grape. Real-time PCR analysis revealed FaCP could be expressed in different strawberry tissues including root, stem, leaf, calyx and fruit, of which the highest expression level in fruit. The FaCP expressed continuously during the whole period of strawberry fruit development and reached the maximum at the pink ripening stage and decreased slightly at the red ripening stage, i ne expression Lewel of FaCP, gradually increased with leaf senescence, which the expression in the young leaves stage （leaf unfolding within 15 days） was low, but in the old leaves （leaf unfolding for more than 45 days）, the FaCP had a higher expression. This result showed that FaCP may play a role in the fruit ripening and leaf senescence.

关 键 词：草莓 半胱氨酸蛋白酶 成熟 基因表达 

分 类 号：S855.3[农业科学—临床兽医学]