海岛棉与陆地棉纤维C4H1基因克隆及表达分析资料.doc

海岛棉与陆地棉纤维C4H1基因克隆及表达分析 曹佳强1，2，吕淑萍2，李波2，杨洋2，张经博2，谢丽霞2，胡文冉2，范玲( （1新疆农业大学农学院乌鲁木齐830052；2新疆农业科学院核所乌鲁木齐830091） 摘要：【目的】比较研究海岛棉与陆地棉纤维不同发育时期C4H1基因的表达和结构差异。【方法】分别从海岛棉和陆地棉纤维中克隆C4H1基因的cDNA全长，并利用DNAMAN、MEGA3.1等软件进行C4H1基因的生物信息学分析；采用半定量和实时荧光定量PCR方法，比较海岛棉和陆地棉不同发育时期的棉纤维中C4H1基因的表达水平。【结果】C4H1基因在海岛棉与陆地棉纤维发育不同时期表达具有差异，海岛棉中该基因的表达高峰期在20 DPA，而陆地棉中该基因的表达高峰期在15 DPA。该基因的核酸序列在两种棉花中存在差异，并导致其编码蛋白的氨基酸序列也存在差异，且海岛棉C4H1蛋白中Thr（苏氨酸）含量高于陆地棉，蛋白等电点也要高于陆地棉。【结论】海岛棉中C4H1基因的表达高峰期晚于陆地棉，两个棉种C4H1蛋白氨基酸序列的差异可能导致其理化性质及生物学功能的改变，该研究为发掘海岛棉纤维品质优异基因奠定了基础。 关键词：陆地棉；海岛棉；基因克隆；纤维品质；基因表达 C4H1 Gene Cloning and Expression Analysis between Sea-island Cotton (Gossypium barbadense L.) Fiber and Upland Cotton (Gossypium hirsutum L.) Fiber CAO Jia-qiang1,2, LV Shu-ping2，LI Bo2, YANG Yang2, ZHANG Jing-bo2, XIE Li-xia2, HU Wen-ran2, FAN Ling2 （1.College of Agronomy, Xinjiang Agricultural University, Urumqi 830052, China；2 Institute of Nuclear and Biological Technologies, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China） Abstract:【Objective】To compare the differences of C4H1 gene expression and structure in different periods of fiber development between sea-island cotton (Gossypium barbadense L.) and upland cotton (Gossypium hirsutum L.)【Method】The full-length cDNA of C4H1 gene was isolated from sea-island cotton and upland cotton fiber respectively. Used DNAMAN, MEGA3.1 software to make bioinformatic analysis of genes. Semi-quantitative interpretation PCR and real-time fluorescence quantitative PCR were used to compare the expression of the target genes in different fiber development periods between two cotton species.【Result】The result shows that C4H1 gene expression was different in different fiber development periods between two cotton species. The expression peak of the C4H1 appeared at 20 DPA in sea-island cotton and 15 DPA in upland cotton. The nucleic acid sequences and coded amino acids of C4H1 were different between two cotton species. The Thr’ (threonine) content and isoelectric point of C4H1 protein in sea-