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杜仲肉桂醇脱氢酶基因全长cDNA克隆及序列分析
引用本文:赵丹,李晓毓,陈建,赵德刚.杜仲肉桂醇脱氢酶基因全长cDNA克隆及序列分析[J].贵州大学学报(农业与生物科学版),2012(4):283-287.
作者姓名:赵丹  李晓毓  陈建  赵德刚
作者单位:[1]贵州大学贵州省农业生物工程重点实验室,贵州贵阳550025 [2]贵州大学生命科学学院,贵州贵阳550025 [3]黄山学院生物与环境科学系,安徽黄山242700 [4]江苏省泗洪县农水局,江苏泗洪223900
基金项目:国家自然科学基金资助(No.30660146和No.30711260268).
摘    要:以杜仲(Eucommia ulmoides Olive)4、5月份新长成的杜仲幼嫩叶片为材料,在克隆一段肉桂醇脱氢酶(cin-namyl alcohol dehydrogenase,CAD)基因的基础上,以杜仲cDNA为模板,采用cDNA末端快速扩增法(Rapid ampli-fication of cDNA Ends,RACE)克隆了5’端828 bp和3’端798 bp cDNA序列,经5’RACE产物和3’RACE产物序列拼接,获得全长为1243bp的杜仲CAD cDNA序列,开放阅读框编码243个氨基酸,命名为EuCAD(GenBank登录号:DQ142643)。与GenBank中序列比对分析发现,该cDNA序列与苹果树、桉树、红橡树中的CAD基因序列同源性均为81%,预测编码的氨基酸序列与苹果树、桉树、红橡树的同源性分别为73%、70%和70%,因此认为是杜仲肉桂醇脱氢酶基因。该基因为首次从杜仲中克隆,为探索木质素的合成调控机理奠定基础。

关 键 词:杜仲  肉桂醇脱氢酶  cDNA末端快速扩增

Cloning and Sequence Analysis of the Full-length cDNA of Cinnamyl Alcohol Dehydrogenase Gene from Eucommia ulmoides Olive
ZHAO Dan,LI Xiao-Yu,CHEN Jian,ZHAO De-Gang.Cloning and Sequence Analysis of the Full-length cDNA of Cinnamyl Alcohol Dehydrogenase Gene from Eucommia ulmoides Olive[J].Journal of Mountain Agriculture & Biology,2012(4):283-287.
Authors:ZHAO Dan  LI Xiao-Yu  CHEN Jian  ZHAO De-Gang
Institution:1. Guizhou Key Labrotary of Agrieultural Bioen- gineering, Guizhou University, Guizhou Guiyang 550025, China; 2. College of Life Sciences, Guizhou Univer- sity, Guizhou Guiyang 550025, China ; 3. Biological and Environmental Sciences, Huangshan University, An- hui Huangshan 242700, China; 4. Department of Agriculture Bureau of Sihong County, Jiangsu Sihong 223900, China)
Abstract:Cinnamyl alcohol dehydrogenase (CAD) plays an important role in the lignin biosynthesis. Cloning and sequence analysis of this gene (CAD) from Eucommia ulraoides Olive were carried out by Rapid Amplifi- cation of cDNA Ends (RACE) in the current work. The sequence analysis showed that the full-length eDNA of CAD contained 1243 bp, whose open reading frame ( ORF ) predicted a protein of 243 amino acids. The eDNA blast in GenBank showed 81% homology with Malus domestica, Eucalyptus gunnii, and Quercus suber, and amino acid blast demonstrated 73% , 70% , 70% homology with that of just-mentioned species, respec- tively, suggesting that full-length eDNA was authentic Eucommia CAD. The Cloning of Eucommia CAD may facilitate to unravel the synthetical mechanism of lignin in plant.
Keywords:Eucommia ulrnoides Olive  CAD gene  RACE
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