两个不同来源的番茄GGPS基因克隆和序列分析  

Cloning and Sequence Analysis of GGPS Gene in Two Sources of Tomato

作　　者：钟秋月[1] 国艳梅[2] 梁燕[1] 王孝宣[2] 杜永臣[2] 朱德蔚[2] 高建昌[2] 戴善书[2] 

机构地区：[1]西北农林科技大学,陕西杨凌712100 [2]中国农业科学院蔬菜花卉研究所,北京100081

出　　处：《华北农学报》2009年第3期15-22,共8页Acta Agriculturae Boreali-Sinica

基　　金：国家重点基础研究发展计划项目(2007CB108800);农业部蔬菜遗传与生理重点开放实验室;中国农业科学院作物科学研究所基金基本科研业务费专项

摘　　要：据本实验室得到的GGPS基因片段与NCBI公布的GGPS基因cDNA(DQ267903)序列设计引物,以两份番茄红素含量差异显著的番茄材料:普通番茄(编号E6203)和源于多毛番茄的渐渗系(编号TA517)为试材,分别克隆其DNA和cDNA序列。序列分析显示:GGPS基因不含有内含子。所推导氨基酸序列比对表明:2个不同来源的GGPS基因存在13个差异位点,其中9个位点的氨基酸性质发生了变化。氨基酸变异导致了2个GGPS基因的二级结构与磷酸化位点的不同,初步推测番茄红素含量的差异可能与氨基酸变异有关;采用双链接头介导的染色体步移技术,克隆了GGPS基因5′侧翼序列,进一步分析结果显示:两份材料GGPS上游区序列差异明显,但所包含的顺式元件的分布相对保守,且存在明显的转录调控序列;利用RT-PCR方法对不同器官的GGPS基因表达进行初步研究,结果表明:GGPS基因主要在果实和花器官中表达。In this study, E6203（common tomato）and TA517（introgression lines of Solanum habrochaites ）which have significant different lyeopene content, were selected to be experimental materials. Specific primers had been designed, which based on GGPS fragment obtained from the subject group of fresh tomato in CAAS and the gene（DQ267903）publicized in NCBI to amplify DNA and cDNA. Sequences analysis indicated that GGPS doesn＇t have intron. Putitive amino acid showed that 13 different residues are existed in the two GGPS ,9 of them characters changed. The mutations lead to the difference of secondary structures, so do phosphorylation sites. It is suggested that the different lyeopene content relat ed with amino acid mutations. 5＇-flanking sequence were obtained by genome walking. Further analysis showed that it contains significant transcriptional regulation sequences and the distribution of cis-acting element was conserved in two GGPS flanking sequence, but sequences had more difference. Semi-quantitative RT-PCR analysis presented that GGPS expressed mainly in fruit and flower.

关 键 词：番茄 GGPS 上游侧翼序列 基因克隆 序列分析 

分 类 号：Q785[生物学—分子生物学]