棉纤维特异表达蓝铜蛋白基因(GhBCP1)的克隆与鉴定  

Isolation and Characterization of a Blue Copper Protein Gene (GhBCP1) Associated with Cotton Fiber Development

作　　者：田琴[1] 李艳军[1] 郭芳[1] 张新宇[1] 王海云[2] 孙杰[1] 

机构地区：[1]石河子大学农学院,新疆兵团绿洲生态农业重点实验室,新疆石河子832003 [2]中国科学院微生物研究所,北京100101

出　　处：《中国农业科学》2010年第3期612-617,共6页Scientia Agricultura Sinica

基　　金：国家“973”计划项目(2004CB117304)

摘　　要：【目的】对从棉纤维细胞分离获得的基因GhBCP1进行序列和表达分析,初步分析其功能。【方法】采用mRNA荧光差异显示结合cDNA末端快速扩增技术克隆基因全长cDNA序列,用生物信息学方法对获得的cDNA序列及推定氨基酸序列进行分析,并用荧光实时定量PCR法研究基因在不同组织中的表达。【结果】克隆了一个棉纤维特异表达基因的全长cDNA,命名为GhBCP1(GenBank登录号:EF222282),该cDNA全长721bp,含有一个编码176个氨基酸蛋白的开放阅读框。BLAST分析表明该基因所编码产物为一个蓝铜蛋白。Southern杂交分析表明该基因在陆地棉(Gossypium hirsytum L.)中有2个拷贝。实时荧光定量PCR分析发现该基因在棉花纤维细胞特异表达,在纤维发育过程中,GhBCP1转录产物的累积主要发生在纤维细胞发育由伸长向次生壁合成转换阶段。【结论】GhBCP1基因的组织特异性和发育阶段性表达初步证明该基因的功能可能与次生壁合成的起始密切相关。[Objective] The aim of this study was to isolate the cDNA sequence of the gene GhBCP1, and to probe the functions of the gene by characterizing it＇s expression patterns in different tissues and developing fibers. [ Method ] The full-length cDNA sequence of GhBCP1 was cloned by Fluorescence differential display （FDD） technique and rapid amplification of the cDNA ends （RACE）. Bioinformatics methods were used to analyze the cDNA and the encoded amino acid sequence, and fluorescence real time quantitative PCR method was used to study the expression pattern of the gene in different tissues. [Result] A full-length cDNA that is specifically expressed in cotton fiber cells was cloned. The gene was designated GhBCP1 （GeneBank accession： EF222282）. The cloned cDNA was 721bp in length which encoded a peptide of 176 amino acids. Blast analysis indicated that this gene encoded a blue copper protein. Southern blot analysis showed that the upland cotton genome contains two copies of the gene. Real-time PCR analysis indicated that GhBCP1 was specifically expressed in fiber cells and its transcripts were more abundant at the transition between the elongation and secondary cell wall synthesis stages. [Conclusion] The results showed that GhBCP1 may play a role at the onset of the secondary cell wall synthesis in developing cotton fiber cells.

关 键 词：棉花 纤维 蓝铜蛋白 特异表达 

分 类 号：S562[农业科学—作物学]