Molecular cloning of dihydroflavonol 4-reductase gene from grape berry and preparation of an anti-DFR polyclonal antibody
文献类型： 外文期刊
作者： Zhang, Ping 1 ; Wen, Peng-Fei 1 ; Wan, Si-Bao 1 ; Wang, Wei 1 ; Pan, Qiu-Hong 1 ; Zhan, Ji-Chen 1 ; Huang, Wei-Dong 1 ;
作者机构： 1.China Agr Univ, Coll Food Sci & Nutr Engn, Beijing 100083, Peoples R China
2.XinJiang Acad Agr Sci, Inst Hort, Urumqi 830000, Peoples R China
3.Shanxi Agr Univ, Coll Hort, Taigu 030801, Shanxi, Peoples R China
关键词： DFR;Escherichia coli expression;antibody..;purification;grape berry
期刊名称：VITIS （ 影响因子：1.339； 五年影响因子：1.306 ）
ISSN： 0042-7500
年卷期： 2008 年 47 卷 3 期
页码：
收录情况： SCI
摘要： Dihydroflavonol 4-reductase (DFR, EC 1.1.1.219) is a key enzyme of the flavonoid pathway, which synthesizes numerous secondary metabolites to determine the quality of grape berry and wine. The full-length dfr cDNA wit h 1014 bp was cloned from grape berry, and then introduced into an expressed plasmid pET-30a (+) vector at the EcoR I and Xho I restriction sites. With induction of the isopropyl-beta-D-thiogalactoside (IPTG), the pET-dfr was highly expressed in Escherichia coli BL21 (DE3) pLysS cells. A fusion protein with the His-Tag was purified through Ni-NTA His Bind Resin and then used as the antigen to immunize a New Zealand rabbit. The resulting antiserum was further purified precipitated by 50 % saturated ammonium sulfate and DEAE-Sepharose FF chromatography to obtain the immunoglobulin G (IgG) fraction. The resulting polyclonal antibody was found capable of immuno-recognizing the DFR of the crude protein extracts from grape berry. This work undoubtedly provides the possibility for further studies on biological regulation of DFR activity in grape berry.
- 相关文献
作者其他论文 更多>>
-
Bioprospecting of a Native Plant Growth-Promoting Bacterium Bacillus cereus B6 for Enhancing Uranium Accumulation by Sudan Grass (Sorghum sudanense (Piper) Stapf)
作者：Wu, Longyuan;Wang, Yanzhi;Sun, Meng;Shi, Chong;Zhang, Lijuan;Wang, Ning;Huang, Wei;Wang, Yanzhi;Sun, Meng;Wang, Wei;Zhang, Lijuan;Zheng, Guofeng
关键词：
Bacillus cereus ; plant growth-promoting activity; uranium stress -
Biocontrol potential of borrelidin metabolites derived from Streptomyces rochei A144 as a fungicide
作者：Wang, Ning;Huang, Wei;Jia, Qiong;Wang, Suling;Wu, Longyuan;Sun, Meng;Wang, Yanzhi;Zhang, Lijuan;Wang, Wei;Song, Bo;Wang, Suling;Wu, Longyuan;Sun, Meng;Wang, Yanzhi;Zhang, Lijuan
关键词：Streptomyces rochei a144; borrelidin; Valsa mali; antifungal activity; biocontrol
-
Assessment of the low lethal effects of emamectin benzoate, chlorantraniliprole, and chlorfenapyr on life parameters and detoxification enzyme activity in Helicoverpa armigera (Lepidoptera: Noctuidae)
作者：Fang, Fengyun;Sun, Hong;Lv, Mingyuan;Chen, Yanlu;Liang, Gemei;Wang, Wei
关键词：Helicoverpa armigera; toxicity; low lethal effect; life table
-
Performing whole-genome association analysis of winter wheat plant height using the 55K chip
作者：Ding, Yindeng;Fan, Guiqiang;Gao, Yonghong;Huang, Tianrong;Zhou, Anding;Fang, Hui;Yu, Shan;Zhao, Lianjia;Shi, Xiaolei;Ding, Sunlei;Hao, Jiahao;Wang, Wei;Song, Jikun;Sun, Na
关键词：winter wheat; plant height; 55K SNP array; GWAS; candidate genes
-
Soybean Sprout Peptides Alleviate Obesity via PI3K-Akt and JAK-STAT Pathway Modulation, Gut Microbiota Regulation, and Metabolic Reprogramming
作者：Wu, Yi;Ling, Yilin;Yang, Yuqi;Long, Haoxing;Kong, Linghao;Song, Peiyang;Lu, Yujie;Zhao, Wanqiu;Wang, Wei;Wu, Yi;Wang, Wei;Zhu, Jingrong;Wang, Nan
关键词：soybean sproutpeptides; obesity; PI3K-Aktpathway; JAK-STAT pathway; gut microbiota; metabolomics; functional food
-
Transcriptomics-Based Study of Immune Genes Associated with Subclinical Mastitis in Bactrian Camels
作者：Ma, Wanpeng;Zhang, Lin;Zhang, Yi;Wang, Yan;Wang, Wei;Liu, Yifan;Zhao, Xueting;Tong, Panpan;Su, Zhanqiang;Ma, Wanpeng;Zhang, Lin;Zhang, Yi;Wang, Yan;Wang, Wei;Liu, Yifan;Zhao, Xueting;Tong, Panpan;Su, Zhanqiang;Yao, Huaibin
关键词：Bactrian camel; mastitis; RNA Seq; immune pathways; gene expression; camel milk
-
Genome-wide association analysis of wheat stem traits using 55K microarrays
作者：Wang, Wei;Zhao, Kai;Sun, Na;Song, JiKun;Fang, Hui;Fan, Guiqiang;Gao, Yonghong;Huang, Tianrong;Ding, Yindeng
关键词：wheat; stem thickness; collapse; GWAS; candidate genes
