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| Construction and Evaluation of a Directed Evolution Library of Catalase |
| TIAN Ya-rong1,2, WANG Ying-hui1,2, XU Mei-ai1,2, LIN Jun1,2, LIN Juan1,2, YE Xiu-yun1,2 |
1. College of Biological Science and Technology, Fuzhou University, Fuzhou 350108,China;
2. National Engineering Laboratory for High-efficient Enzyme Expression, Fuzhou 350108, China |
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Abstract The directed evolution of wild-type catalase from Cervus nippon was studied, which mimicked evolution on a laboratory timescale. The mutant library of catalase gene was constructed using error-prone PCR and DNA shuffling, and its sequencing result showed mutant rate to be 0.329% and 27.58%. The mutant rate using error-prone PCR was 10 fold higher than normal PCR, while the mutant rate using DNA shuffling was much more higher. However, tremendous mutant rate may affect or even disable gene transcription and translation. A suitable mutant rate is needed to guarantee fluent gene expression. Furthermore, both wild-type and mutant catalase sequences selected randomly were analysed to obtain their Neighbor-Joining phylogenetic trees.
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Received: 25 April 2011
Published: 25 September 2011
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