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| Genetic Analysis of T-DNA Insertion Site and Characterization of One Aspergillus niger Mutant with Low Cellulase Activity |
| HUANG Wen, YANG Hong-jiang, QIN Hui-bin |
| Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China |
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Abstract Selective medium with sodium carboxymethyl cellulose (CMC) as only carbon source was employed for screening T-DNA insertional library of Aspergillus niger, from which mutant AN-108 with low cellulase activity was isolated. Its enzyme activity is 83.3% to that of wild type Aspergillus niger strain. Solid state fermentation analysis showed that no cellulase activity difference was obtained between mutant AN-108 and wild type strain of A. niger. It’s hypothesized that sugars existed in solid state fermentation media might contribute to the restoration of cellulase activity of the mutant. To test this possibility, CMC media with different sugars were used for culturing mutant AN-108. Hydrolyze zones around colonies were observed in all plates with sugars. The experiment result suggested that sugar might function as expression inducers overcoming the mutation effect. To identify the gene disrupted in mutant AN-108, sequence analysis of T-DNA insertion site was performed using inverse PCR method. The result of sequence blast indicated that the disrupted gene had 90% identities to the gene An14g03730 of A. niger, which encoded a proline-rich protein.
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Received: 08 November 2010
Published: 26 April 2011
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