两个不同来源的番茄GGPS基因克隆和序列分析

doi:10.7668/hbnxb.2009.03.004

华北农学报 ›› 2009, Vol. 24 ›› Issue (3): 15-22. doi: 10.7668/hbnxb.2009.03.004

所属专题： 番茄；生物技术；

两个不同来源的番茄GGPS基因克隆和序列分析

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

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

收稿日期:2009-02-27 出版日期:2009-06-28
通讯作者: 杜永臣(1955 - ),男,河北昌黎人,研究员,博士,主要从事蔬菜遗传育种研究。
作者简介:钟秋月(1982-), 女, 河北保定人, 硕士, 主要从事蔬菜育种与生物技术研究.
基金资助:
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

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

ZHoNG Qiu yue¹, GUo Yan mei², LIANG Yan¹, WANG Xiaoxuan², DU Yong chen², ZHU Dewei², GAo Jian chang², DAI Shan shu²

1. Northwest Agriculture and Forestry Universiry,Yangling 712100,China;
2. Institute of Vegetables and Flowers,Chinese Academy of Agriculture Sciences,Beijing 100081,China

Received:2009-02-27 Published:2009-06-28

摘要/Abstract

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TASl7)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-PER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.

关键词: 番茄, GGPS, 上游侧翼序列, 基因克隆, 序列分析

Abstract: In this study,E6203(common tomato)and TA517(introgression lines of Solanum habrochaites)which have significant different lycopene 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)publi2 cized 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 lycopene content relat2 ed with amino acid mutations. 5′ 2flanking sequence were obtained by genome walking. Further analysis showed that it con2 tains significant transcriptional regulation sequences and the distribution of cis2acting element was conserved in two GGPS flanking sequence,but sequences had more difference. Semi2quantitative RT2PCR analysis presented that GGPS expressed mainly in fruit and flower.

Key words: Tomato, GGPS, 5&prime, 2flanking regions, Gene cloning, Sequence analysis

中图分类号:

Q785

钟秋月, 国艳梅, 梁燕, 王孝宣, 杜永臣, 朱德蔚, 高建昌, 戴善书. 两个不同来源的番茄GGPS基因克隆和序列分析[J]. 华北农学报, 2009, 24(3): 15-22. doi: 10.7668/hbnxb.2009.03.004.

ZHoNG Qiu yue, GUo Yan mei, LIANG Yan, WANG Xiaoxuan, DU Yong chen, ZHU Dewei, GAo Jian chang, DAI Shan shu. Cloning and Sequence Analysis of GGPS Gene in Two Sources of Tomato[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(3): 15-22. doi: 10.7668/hbnxb.2009.03.004.

http://www.hbnxb.net/CN/Y2009/V24/I3/15

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