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Effects of nitrogen ion irradiation on endoglucanase activity and gene mutation of Bacillus subtilis Bac01
Authors:L Jie  MAO Peihong  JIN Xiang  YU Long  YING Hanjie
Affiliation:1. College of Biotechnology and Pharmaceutical Engineering,Nanjing University of Technology,210009 Nanjing,China;Center of Ion Beam Biotechnology,College of Physics Science and Technology,Xinjiang University,830008 Urumqi,China
2. Center of Ion Beam Biotechnology,College of Physics Science and Technology,Xinjiang University,830008 Urumqi,China
3. College of Biotechnology and Pharmaceutical Engineering,Nanjing University of Technology,210009 Nanjing,China
Abstract:Bacillus subtilis Bac01 was mutated by 15 keV N+ ions of 1.5×1016 cm-2.The mutant strain Bacll with high yield of endoglucanase was isolated using carboxymethylcellulose sodium and congo red indicative plates.It exhibited higher endoglucanase activitiy (381.89IU) than the original strain Bac01 (93.33IU).Two 1,500 bp endoglucanase gene fragments were obtained with PCR amplification from B.subtilis Bac01 and mutant strain Bac11.BLAST comparison result indicated that 10 nucleotides mutated.Bioinformatics methods were used to analyze the two predicted amino acid sequences,and it was found that 5 amino acid residues changed,being all in the cellulose-binding domain of endoglucanase.
Keywords:Ion irradiation  Bacillus subtilis  Endo-1  Activity  Gene mutation
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