刺五加多向耐药性转运蛋白基因的克隆及其表达对皂苷含量的影响  

Cloning of pleiotropic drug resistance transporter gene from Eleutherococcus senticosus and effect of its expression on saponins content

作　　者：邢朝斌[1] 孟春燕[2] 修乐山[1] 劳凤云[3] 庄鹏宇[3] 柴丽花[1] 

机构地区：[1]河北联合大学生命科学学院,河北唐山063000 [2]河北联合大学公共卫生学院,河北唐山063000 [3]河北联合大学药学院,河北唐山063000

出　　处：《浙江农业学报》2014年第2期351-355,共5页Acta Agriculturae Zhejiangensis

基　　金：国家自然科学基金项目(30701086);河北省自然科学基金项目(C2009001252);河北省自然科学基金-石药集团医药联合研究基金(H2012401006);河北联合大学培育基金(GP201306)

摘　　要：利用简并引物进行聚合酶链式反应(PCR)扩增获得长度为693 bp的刺五加(Eleutherococcus senticosus)多向耐药性(pleiotropic drug resistance,PDR)转运蛋白基因,GenBank登录号为KC473536,其编码的231个氨基酸的蛋白属于PDR转运蛋白的核苷酸结合结构域。氨基酸序列与毛果杨(Populus trichocarpa)、蓖麻(Ricinus communis)和水稻(Oryza sativa)的PDR氨基酸序列一致性分别为88.74%,90.04%和88.31%。RT-PCR法检测PDR在刺五加不同生长发育时期和不同器官中表达情况和分光光度法测定刺五加总皂苷含量的结果显示:刺五加PDR基因在整个生长期中均有表达,与皂苷在整个生长期中均有合成的特点相符,但两者的相关性未达显著水平。PDR基因在叶片和叶柄中高表达的特点与刺五加皂苷仅存在于叶中的特点相符,两者间存在显著的正相关关系(P<0.05)。cDNA encoding pleiotropic drug resistance （PDR） gene, whose length was 693 bp, was amplified from Eleutherococcus senticosus,by means of polymerase chain reaction （PCR） with degenerate oligo nucleotide primers , the gene has been deposited in GenBank under accession number KC 473536.It encoded a protein of 231 amino acids belonging to PDR＆#39;s nucleotide binding domain.In comparison with E.senticosus PDR with those of Populus trichocarpa, Ricinus communis and Oryza sativa on the amino acid sequence, the amino acid homology was 88.74%, 90.04% and 88.31%.The expression of PDR in different organs of E.senticosus in various growing periods was detected by RT-PCR, and E.senticosus saponins content was measured by spectrophotometry method , the results showed that PDR gene expressed in the whole growing periods of E.senticosus, which was consistent with the＆amp;nbsp;characteristic of saponins formation in the whole growing periods , and the correlation of the two was not significant . The characteristic of high expression of PDR gene in leaf and petiole was consistent with the phenomenon that saponins were only found in leaf , which showed significantly positive correlation （P〈0.05）.

关 键 词：刺五加 多向耐药性转运蛋白 克隆 表达分析 皂苷 

分 类 号：S567[农业科学—中草药栽培]