刺五加法尼基焦磷酸合酶基因的克隆、 生物信息学及表达分析

目的:克隆刺五加的法尼基焦磷酸合酶(farnesyl diphosphate synthase,FPS)基因,并对其进行生物信息学和表达分析.方法:采用cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)技术克隆刺五加FPS基因的全长cDNA序列.运用生物信息学方法对该基因进行分析,预测其编码蛋白的结构与功能.并通过RT-PCR法检测FPS在不同器官中的表达情况.结果:刺五加FPS基因的cDNA全长为1 499 bp,开放阅读框长1 029 bp,编码342个氨基酸的蛋白,包含2个富含天冬氨酸(DDXXD)的保守功能域.FPS蛋白无跨膜区域,定位于细胞质中.RT-PCR的结果显示,刺五加FPS基因在各器官中均有表达,但表达最具有显著差异(P＜0.05).结论:首次分离并报道了刺五加FPS的cDNA克隆,并证实其在不同器官中的表达量不同,为进一步研究FPS对刺五加皂苷合成的影响和表达调控奠定了基础.

Molecular cloning of farnesyl diphosphate synthase from Eleutherococcus senticosus and its bioinformatics and expression analysis

Objective: To clone farnesyl diphosphate synthase (FPS) gene from Eleutherococcus senticosus and analyze the bioinformatics and expression of the gene. Method: The FPS full length cDNA was cloned by rapid amplification of cDNA ends (RACE). The data was analyzed by bioinformatics method,the structure and function of FPS was deduced. The expression of FPS in different organ of E. senticosus was detected by RT-PCR. Result: The full length of FPS cDNA was 1 499 bp containing a 1 029 bp ORF that encoded 342 amino acids. The deduced protein sequence exhibited two Asp riches conserved motifs (DDXXD). Without transmembrane domain,FPS was located in cytoplasm. RT-PCR result showed that FPS gene expressed in different organs of E. senticosus. The expression amounts of FPS in different organs were different significantly (P<0.05). Conclusion: The FPS gene of E. senticosus was successfully cloned for the first time,and provided a stable foundation for studying on its effect and expression control on E. senticosus saponins biosynthesis.