摘要
以五年生人参根组织为试验材料,克隆得到人参法尼基焦磷酸合成酶(FPS)基因。通过生物信息技术对其氨基酸序列进行比对,使用MEGA5.1构建人参与相关物种的FPS系统进化树,运用实时荧光定量PCR方法检测FPS基因在不同生长时期的表达量,用高效液相色谱法测定根中8种单体皂苷含量,进行相关性分析。结果显示:人参FPS基因c DNA大小为1 134 bp,含有一个开放阅读框,编码342个氨基酸。FPS编码蛋白相对分子质量为39 567,等电点为5.83,不含信号肽,无跨膜区。二级结构中主要为α-螺旋,占58.19%。同源性分析显示,FPS基因与西洋参(ADJ68004)、三七(AGS79228)核苷酸序列相似性为99%。实时荧光定量PCR结果显示,FPS基因在人参各生长时期中,以展叶期表达量最高。相关性分析表明,FPS基因的表达与Rb3含量显著正相关(P<0.05),与三醇型皂苷含量呈正相关,但相关性不显著。
Using five-year ginseng root tissue as experimental materials,ginseng farnesyl pyrophosphate synthase（ FPS） gene was cloned. Protein characteristics were analyzed by bioinformatics,and phylogenetic tree of FPS was constructed using MEGA5. 1. Tissue expression in different growth stages was analyzed using RT-PCR technology. Content of 8 kinds of ginsenosides in root was determined by HPLC and correlation analysis was conducted.1 134 bp FPS sequence was obtained,which has a complete open reading frame,encoding an unstable protein with 342 amino acids. FPS encoding protein has isoelectric point of 5. 83 and a calculated relative molecular weight about 39 567,without transmembrane regions and signal peptide. In the secondary structure,the main alpha helix accounted for 58. 19%. The homologous analysis indicated nucleotide sequence was 99% homologous with Panax quinquefolius D S（ ADJ68004） and Panax notoginseng D S（ AGS79228）. Real-time quantitative PCR showed that FPS gene was expressed in all growth stages. Expression level of Panax ginseng FPS was the highest in leaf-expanding period.Correlation analysis showed that expression of FPS gene was significantly positively correlated with content of Rb3（P0.05）,and positively correlated with content of panaxtrolsaponins,but the correlation was not significant.
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2017年第6期695-702,708,共9页
Journal of Jilin Agricultural University
基金
国家中药材产业技术体系岗位科学家专项(CARS-21)
国家自然科学基金项目(31270371)
关键词
人参
法尼基焦磷酸合成酶
生物信息技术
人参皂苷
相对表达量
相关性
Panax ginseng
farnesyl diphosphate synthase
biological information technology
gin-senoside
relative quantitative expression
correlation
