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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 |
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| 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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