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小麦返白系胆色素原脱氨酶纯化及编码cDNA序列分析
引用本文:程冬梅,范三红,刘香莉,陈根云,邓志勇,郭蔼光.小麦返白系胆色素原脱氨酶纯化及编码cDNA序列分析[J].中国生物化学与分子生物学报,2006,22(12):973-978.
作者姓名:程冬梅  范三红  刘香莉  陈根云  邓志勇  郭蔼光
作者单位:1. 西北农林科技大学陕西省农业分子生物学重点实验室,杨凌,712100;徐州师范大学生命科学学院,江苏,徐州,221116
2. 西北农林科技大学陕西省农业分子生物学重点实验室,杨凌,712100
3. 中国科学院上海植物生理生态研究所,上海,200031
基金项目:国家自然科学基金;高等学校博士学科点专项科研项目
摘    要:以小麦(Triticum aestivum)返白系F772为材料,通过粗提、加热处理、硫酸铵分级沉淀和凝胶柱层析等方法,对胆色素原脱氨酶(PBGD)进行了提取纯化,纯化倍数为1092,得率为15%.纯化的PBGD约为37 kD.对其部分生化性质的研究表明:高浓度的NH+4对酶活性有强烈的抑制,光照处理可以使酶活性降低.对纯化后的PBGD N末端氨基酸序列进行了测定.根据N端序列设计简并引物,获得了PBGD的cDNA全部编码区序列.它编码一个351个氨基酸的前体蛋白,有一个叶绿体导肽的序列.通过比较小麦PBGD与来自与其他物种的同源蛋白表明,有些氨基酸残基非常保守.

关 键 词:小麦返白系  胆色素原脱氨酶  N末端氨基酸测序  纯化  cDNA克隆  
收稿时间:2006-5-29
修稿时间:2006年5月29日

Purification and Sequence Analysis of cDNA Coding Region for Porphobilinogen Deaminase from a Stage Albinism Line of Wheat
CHENG Dong-Mei,FAN San-Hong,LIU Xiang-Li,CHEN Gen-Yun,DENG Zhi-Yong,GUO Ai-Guang.Purification and Sequence Analysis of cDNA Coding Region for Porphobilinogen Deaminase from a Stage Albinism Line of Wheat[J].Chinese Journal of Biochemistry and Molecular Biology,2006,22(12):973-978.
Authors:CHENG Dong-Mei  FAN San-Hong  LIU Xiang-Li  CHEN Gen-Yun  DENG Zhi-Yong  GUO Ai-Guang
Institution:1)Shaanxi Key Laboratory of Agricultural Molecular Biology, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100, Shaanxi, China; 2) College of Life Sciences, Xuzhou Normal University, Xuzhou 221116, Jiangsu, China;3) Institute of Plant Physiology and Ecology, CAS, Shanghai 200031, China
Abstract:Porphobilinogen deaminase (PBGD) was purified from F772, a mutant of ‘the stage albinism line of wheat’, following the procedures including crude extracts, heat treatment, ammonium sulfate fraction and column chromatography. The enzyme was purified 1 092 fold with a special activity recovery of 15%. The wheat BGD is a 37 kD protein. The activity of enzyme could be greatly inhibited by high contents of ammonium ion as well as lighting treatment. The N-terminal amino acid sequence was determined for PBGD, based on which, PBGD cDNA was obtained with the degenerate primers. Sequence analysis of coding region of PBGD indicated that it encodes a precursor of 351 residues with a typical chloroplast-targeted peptide. In comparison of wheat PBGD and its homologs from other species, it was found that there are some highly conserved amino acid residues among these proteins.
Keywords:the stage albinism line of wheat  porphobilinogen deaminase  N-terminal amino acid sequencing  purification  cDNA cloning
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