根据棉花GhCCR1基因的cDNA序列设计引物,采用PCR技术从棉花中克隆了GhCCR1基因的DNA序列,并采用半定量RT-PCR方法分析了GhCCR1基因在不同发育阶段棉纤维中的表达情况.结果表明:GhCCR1编码区DNA序列长度为1 161 bp,包含4个外显子和3个内含子,内含子富含AT,所有外显子/内含子交接点都遵从gt/ag剪接规则.半定量RT-PCR检测表明,GhCCR1基因在不同发育阶段的棉纤维中均有表达,在开花后20 d的棉纤维中表达量最高,说明该基因可能参与调控棉纤维细胞的伸长和次生壁的增厚过程.
Primers were designed according to the cDNA sequence of cotton GhCCR1.PCR method was used to clone GhCCR1 from cotton and the expression of GhCCR1 in fibers at different developmental stages was analyzed by semi-quantitative RT-PCR.A length of 1 161 bp from genomic DNA of GhCCR1 was cloned by PCR.The genomic DNA of GhCCR1 contains four exons and three introns.Analysis of these introns indicated that they were rich in AT.All exon/intron junctions of the gene contain the gt/ag consensus splicing site.Semi-quantitative RT-PCR analysis revealed that GhCCR1 was expressed during different fiber developmental stages,and reached to the peak of expression in 20 days post anthers(DPA) fiber cells,suggesting that GhCCR1 may be involved in regulating fiber elongation and secondary wall thickening.