农业生物技术学报2013，Vol.21Issue(2)：158-164,7.DOI:10.3969/j.issn.1674-7968.2013.02.004
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)
摘要
Abstract
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. The expression level of FaCP gene in leaves was 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Key words
Strawberry(Fragaria×ananassa), Cysteine protease, Fruit ripening, Gene expression引用本文复制引用
朱海生,陈敏氡,温庆放,林珲..草莓半胱氨酸蛋白酶基因(FaCP)的克隆及表达分析[J].农业生物技术学报,2013,21(2):158-164,7.基金项目
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)和福建省农业科学院科技创新团队重点科研项目(No.CXTD2011-20) （No.31101534）
