论文部分内容阅读
目的对杯萼海桑Sonneratia alba萜类化合物生物合成途径的关键酶法呢基焦磷酸合成酶(Farnesyl pyrophosphate synthase,FPS)基因的编码cDNA序列进行克隆,为研究杯萼海桑萜类化合物生物合成与基因调控奠定基础。方法结合杯萼海桑根的转录组注释,根据编码区序列设计引物,通过PCR方法克隆杯萼海桑FPS(SaFPS)基因的编码区cDNA。结果PCR扩增了一个长1 029 bp的基因片段,该片段编码由342个氨基酸组成的SaFPS。同源性比对结果显示基因编码蛋白与甘草的FPS具有氨基酸一致性达86%。其具有异戊烯基转移酶的2个典型保守功能域。进化树分析结果显示,与甘草Glycyrrhiza uralensis、苜蓿Medicago truncatula、羽扇豆Lupinus albu具有较近的亲缘关系。实时荧光定量PCR结果显示SaFPS基因在花中表达量较高,果实、茎、叶中表达量相对较低。结论首次从红树林植物杯萼海桑中克隆FPS基因获得其编码区序列,SaFPS基因具有组织表达特异性。本研究为分析基因表达特性及其在生物合成中的功能奠定基础。
Objective To clone the cDNA sequence of the key enzyme, Farnesyl pyrophosphate synthase (FPS) gene in Sonneratia alba ternary biosynthesis pathway, Lay the foundation with gene regulation. Methods According to the transcriptome annotation of Calyx japonicus, primers were designed according to the sequence of coding region, and the coding region cDNA of FPS (SaFPS) gene was cloned by PCR. Results A gene fragment of 1 029 bp was amplified by PCR. This fragment encoded a 342 amino acid SaFPS. Homology comparison showed that the amino acid identity of gene encoding protein and licorice FPS was 86%. It has two typical conserved domains of prenyltransferase. The results of phylogenetic tree analysis showed that it had close genetic relationship with Glycyrrhiza uralensis, Medicago truncatula and Lupinus albu. Real-time PCR results showed that the expression level of SaFPS gene in flowers was high, and the expression level of fruit, stem and leaf was relatively low. Conclusion For the first time, the coding region sequence of FPS gene was cloned from mangrove Cupped Cuprophyllidae, and the SaFPS gene has the specificity of tissue expression. This study laid the foundation for the analysis of gene expression characteristics and its function in biosynthesis.