利用mRNA荧光差异显示技术规模化筛选棉纤维特异表达基因

Identification of Genes that are Specifically/preferentially Expressed in Developing Cotton Fibers by mRNA Fluorescence Differential Display (FDD)

以陆地棉 (GossypiumhirsutumL .)品种TM 1开花后 9d、2 1d、2 7d三个不同发育时期的棉花纤维为材料 ,利用mRNA荧光差异显示 (FDD)技术 ,筛选到 10 9个差异显示的cDNA片段。在此基础上 ,结合两轮反Northern杂交筛选和Northern杂交分析 ,获得了多个仅在棉花纤维细胞中特异表达或在纤维中优先表达基因的cDNA片段 ,序列测定和数据库搜索分析表明 ,这些cDNA片段中的多数还未有报道。本工作为克隆上述基因的全长cDNA ,并进一步研究它们在棉纤维发育中的功能奠定了基础。

Fluorescence differential display (FDD) technique was used to identify genes that are specifically or preferentially expressed in different developmental stages of cotton fiber cells. One hundred and nine differentially displayed cDNA fragments were isolated using 9, 21 and 27 DPA (days postanthesis) fibers as experimental materials. By a combination of two rounds of reverse Northern hybridization and Northern blot analyses, a number of such cDNA fragments were proved to represent fiber-specific/preferential genes. Sequencing determination and database searching indicated that most of these genes are novel. This work is an important step towards cloning the full-length cDNAs and characterizing the cellular functions of aforementioned genes in fiber development.