棉花中两个新的F-box蛋白基因的克隆与表达分析

Cloning and Expression Analysis of Two New Genes Encoding F-box Proteins in Cotton (Gossypium hirsutum L. )

以棉花EST序列分析为基础,利用电子克隆和RT-PCR技术,从棉花中克隆了2个新的F-box蛋白基因,分别命名为GhFB1(GenBank核酸数据库登录号:EF419428)和GhFB2(GenBank登录号:EF503623)。GhFB1cDNA全长866bp,编码162个氨基酸;GhFB2cDNA全长657bp,编码161个氨基酸,而且两者均含有植物F-box蛋白家族特有的F-box结构域。序列比较分析表明,GhFB1和GhFB2蛋白与水稻OsGID2,拟南芥AtSLY1具有高同源性,是棉花中F-box蛋白家族的新成员。RT-PCR结果表明,GhFB1和GhFB2在根、下胚轴、叶、茎、花和纤维中都有表达,而且均在棉花纤维起始和伸长时期有优势表达,推测这2个基因在棉纤维早期发育中可能起重要作用。

In the attempt to elucidate the pivotal function of GA signaling in the initiation and elongation of cotton fiber, two new F-box protein genes were isolated from Gossypiurn hirsutum for the first time using electronic cloning and RT-PCR methods. They were named GhFB1 and GhFB2, respectively and were submitted to NCBI and the GenBank accession were EF419428 and EF503623. The cDNA of GhFB1 contained 866 bp and encoded a putative protein of 162 aa, while GhFB2 was 920 bp in length, encoding a predicted protein of 161 aa. The encoding proteins have the typical F-box motif of F-box protein family. Bioinformatics analysis indicated that the identities of the deduced GhFB1 and GhFB2 amino acid with Oryza sativa protein, OsGID2, and Arabidopsis thaliana protein, AtSLY1, are very high. Sequence analysis indicated that these two genes are new members of the F-box protein gene family. The expression patterns of GhFB1 and GhFB2 was analyzed by RT-PCR in cotton. It was demonstrated that these two F-box protein genes were both expressed in root, hypocotyls, leaf, stem and fibers, and the highest level of transcripts was to accumulate in the fibers at the stage of initiation and elongation, suggesting that the two F-box genes, GhFB1 and GhFB2, might play important roles in the early stage of fiber development.