海岛棉GbHCT基因的表达分析

Expression Analysis of GbHCTGene in Gossypium barbadence L.

通过海岛棉基因组数据库和GSDS 2.0在线生物信息学软件,分析GbHCT基因的结构,利用实时荧光定量PCR技术,检测GbHCT基因在海岛棉不同组织和棉纤维不同发育时间的表达量。结果表明,该基因DNA序列长度为1 953bp,开放阅读框长度为1 311bp,编码436个氨基酸,在海岛棉基因组中对应scaffold3453序列,含有1个内含子和2个外显子。GbHCT基因在不同组织和纤维发育不同时期中均有表达,在各个组织中,苞叶和花中表达量最高,在棉纤维不同发育时期开花后5d和25d表达量最强。该基因可能参与棉纤维发育伸长期和次生壁增厚期。为深入研究该基因的功能,构建植物表达载体pEGAD-GbHCT并转入根癌农杆菌,为下一步研究奠定试验基础。

In this study,the structure of gene GbHCT was analyzed by using an online bioinformatics software genomic database of Gossypium barbadence and GSDS 2.0.The expression of GbHCTgene in different tissues and different fiber development stages of cotton was analyzed by real-time quantitative PCR.A length of 1 953 bp from genomic DNA of GbHCT was cloned by PCR.GbHCTcontain open reading frames of 1 311 bp encoding proteins of 436 amino acid residues.The genomic DNA of GbHCT contained two exons and one intron which were located in the scaffold 3 453 of Gossypium barbandense L.Real-time quantitative PCR analysis revealed that GbHCT was expressed at higher level in bracts and flowers,and it was predominantly expressed in 5dand 25 dduring cotton fiber development,so we suggested that this gene may be involved in regulating fibre elongation and secondary wall thickening.Plant expression vector pEGAD-GbHCT was constructed and transferred into the Agrobacterium tumefaciens EHA105 for further experiment.