吴茱萸SOD基因片段克隆和SNP分析  

Cloning of SOD Gene Segments and SNP Analysis in Evodia rutaecarpa

作　　者：吴波[1] 高丹[2] 潘超美[3] 罗光明[2] 张寿文[2] 

机构地区：[1]江西中医学院江西省中药种质资源工程技术研究中心,江西南昌330004 [2]江西中医学院药学院,江西南昌330004 [3]广州中医药大学中药学院,广东广州510006

出　　处：《江西农业大学学报》2011年第6期1206-1211,共6页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家"十二五"科技支撑计划项目(2011BAI04B04)

摘　　要：根据GenBank已报道的多种植物Cu/Zn-SOD基因和Mn/Fe-SOD基因的保守区域,分别设计了两对用于克隆10种吴茱萸属植物Cu/Zn-SOD基因和Mn/Fe-SOD基因核心片段的简并引物。试验所得的Cu/Zn-SOD基因片段长度为375 bp,Mn/Fe-SOD基因片段长度为429 bp,分别编码125个和143个氨基酸残基。使用DAM-BE和DNAStar5.0生物统计软件对序列进行同源性比对,10种吴茱萸属植物与其他多种植物的Cu/Zn-SOD基因核苷酸序列同源性大于74%,氨基酸序列同源性大于79%;Mn/Fe-SOD基因的核苷酸序列同源性大于78%,氨基酸序列同源性大于81%。在所克隆的Cu/Zn-SOD基因片段中有21个核苷酸SNPs位点,导致6个氨基酸位点变异,多态性频率为1SNP/17.9 bp,核苷酸和氨基酸变异度分别为5.6%和4.8%;在所克隆的Mn/Fe-SOD基因片段中有17个核苷酸SNPs位点,导致3个氨基酸位点变异,多态性频率为1SNP/25.2 bp,核苷酸和氨基酸变异度分别为4.0%和2.1%。采用Neighbour-Joining聚类法,对10种吴茱萸属植物的Cu/Zn-SOD基因片段的核苷酸序列进行聚类分析,构建了精确遗传聚类图谱。Two pairs of degenerate primers were designed according to the two conserved regions on the amino acid sequences of Cu/Zn-SOD gene and Mn/Fe-SOD gene from kinds of plants reported in GenBank in order to clone the core fragments of these two genes in ten Evodia rutaecarpa＇s plants.The length of Cu/Zn-SOD gene and Mn/Fe-SOD gene were 375 bp and 429 bp respectively.Every nucleotide sequence of Cu/Zn-SOD gene and Mn/Fe-SOD gene encoded 125 amino acids and 143 amino acids respectively.The biology softwares of DAMBE and DNA Star5.0 were used to analyze these sequences.The nucleotide similarity with other plants was above 74% and the amino acid similarity was above 79% at ten Cu/Zn-SOD gene fragments than other plants.The nucleotide similarity was above 78% and the amino acid similarity was above 81% in ten Mn/Fe-SOD gene fragments.Twenty-one sites of single nucleotides were discovered in the ten Cu/Zn-SOD gene fragments.Six amino acids sites transformed because of twenty-one single nucleotides variation.Seventeen variation sites of single nucleotides were discovered in the ten Mn / Fe-SOD gene fragments.Three amino acid sites transformed because of the variation of seventeen single nucleotides.Accoring to the calculation result,the polymorphism frequences in Cu / Zn-SOD gene and Mn / Fe-SOD gene were 1SNP /17.9 bp and 1SNP /25.2 bp,the nucleotide degrees of variation were 5.6% and 4.0%,the amino acid degrees of variation were 4.8% and 2.1%,respectively.Genetic diversity of cloned ten Cu / Zn-SOD gene fragments was analyzed by using the clustering procedure of Neighbor-Joining.

关 键 词：吴茱萸 SOD 基因克隆 SNP 

分 类 号：Q943.2[生物学—植物学]