Purification and Sequence Analysis of cDNA Coding Region for Porphobilinogen Deaminase from a Stage Albinism Line of Wheat
CHENG Dong-Mei 1),2)**, FAN San-Hong 1)**, LIU Xiang-Li 1)，CHEN Gen-Yun3), DENG Zhi-Yong 1), GUO Ai-Guang 1)*
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.
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.
CHENGDong-Mei,,FANSan-Hong,LIUXiang-Li;CHENGen-Yun;DENGZhi-Yong;GUOAi-Guang. Purification and Sequence Analysis of cDNA Coding Region for Porphobilinogen Deaminase from a Stage Albinism Line of Wheat. Chinese Journal of Biochemistry and Molecular Biol, 2006, 22(12): 973-978.
