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| Cloning and analysis of a γ-tocopherol methyltransferase gene from Brassica oleracea and the function of its recombinant protein | |
| 作者姓名： | OUYANG Qing FAN Chuntao SUN Hui ZHANG Yuman BAI Shuangyi CAI Wenqi |
| 作者单位： | Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China,College of Life Sciences, Beijing Normal University, Beijing 100875, China,Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China,Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China,Department of Landscape Architecture, China Agricultural University, Beijing 100094, China,Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China |
| 基金项目： | Supported by the National Natural Science Foundation of China (Grant No. 30270856) |
| 摘 要： | The full-length cDNA (BoTMT) of γ-TMT is obtained from Brassica oleracea by 3′- and 5′-RACE methods. The 1265 bp cDNA contains an open reading frame of 1044 bp, which encodes a protein of 347 amino acids with a predicted chloroplast transit peptide and two S-adenosylmethionine (SAM)-binding domains. Sequence analysis shows that the deduced protein shares 41.8%～86.5% similarity to known γ-TMTs. Semi-quantitative RT-PCR reveals that BoTMT is expressed preferentially in flowers and leaves of B. oleracea. The recombinant γ-TMT protein is obtained by cloning its encoding region into the prokaryotic expression vector pET30a. The protein expressed in E. coli accounts for 22% of total bacterial protein. The enzyme activity assay indicates that the recombinant protein has relatively high activity to convert γ-tocopherol to α-tocopherol in vitro. |
| 关 键 词： | Brassica oleracea gamma-tocopherol methyltransferase gene and expression enzyme activity assay t |
Cloning and analysis of a γ-tocopherol methyltransferase gene from Brassica oleracea and the function of its recombinant protein |
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| OUYANG Qing,FAN Chuntao,SUN Hui,ZHANG Yuman,BAI Shuangyi,CAI Wenqi.Cloning and analysis of a γ-tocopherol methyltransferase gene from Brassica oleracea and the function of its recombinant protein[J].Progress in Natural Science,2003,13(7):511-516. | |
| Authors: | OUYANG Qing FAN Chuntao SUN Hui ZHANG Yuman BAI Shuangyi CAI Wenqi |
| Institution: | 1. Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China 2. College of Life Sciences, Beijing Normal University, Beijing 100875, China 3. Department of Landscape Architecture, China Agricultural University, Beijing 100094, China |
| Abstract: | The full-length cDNA (BoTMT) of γ-TMT is obtained from Brassica oleracea by 3′- and 5′-RACE methods. The 1265 bp cDNA contains an open reading frame of 1044 bp, which encodes a protein of 347 amino acids with a predicted chloroplast transit peptide and two S-adenosylmethionine (SAM)-binding domains. Sequence analysis shows that the deduced protein shares 41.8%～86.5% similarity to known γ-TMTs. Semi-quantitative RT-PCR reveals that BoTMT is expressed preferentially in flowers and leaves of B. oleracea. The recombinant γ-TMT protein is obtained by cloning its encoding region into the prokaryotic expression vector pET30a. The protein expressed in E. coli accounts for 22% of total bacterial protein. The enzyme activity assay indicates that the recombinant protein has relatively high activity to convert γ-tocopherol to α-tocopherol in vitro. |
| Keywords: | Brassica oleracea gamma-tocopherol methyltransferase gene and expression enzyme activity assay t |
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