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.
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
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.
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