Deletion Analysis of GH7 Endoglucanase Gene (cel7B) Promoter Region in a Talaromyces cellulolyticus ligD-Disrupted Strain.

Tatsuya Fujii, Hiroyuki Inoue, Kazuhiko Ishikawa, Tamotsu Hoshino

Applied biochemistry and biotechnology 2017 Dec

filter terms:

Talaromyces cellulolyticus is expected to become an industrial cellulase producer. In this study, we performed deletion analysis of the promoter region of the GH7 endoglucanase gene (cel7B), which encodes one of the major cellulases, using a β-glucuronidase reporter system. To obtain strains that harbor each gene cassette at the same locus, we had to improve the homologous recombination frequency. Hence, the ligD gene, encoding DNA ligase IV, was disrupted by homologous recombination. After that, the introduced pyrF marker gene, encoding orotate phosphoribosyl transferase, was deleted by a marker recycling system. The resultant strain, YDLP, exhibits high homologous recombination frequency. These data suggest that this approach will drastically improve the genetic modification tools of T. cellulolyticus. We obtained 7 strains for reporter analysis using YDLP as the host strain. Reporter analysis revealed that the promoter region between -812 and -612 is important for expression of cel7B. These results imply a relationship between this region and novel transcriptional factors.

Citation

Tatsuya Fujii, Hiroyuki Inoue, Kazuhiko Ishikawa, Tamotsu Hoshino. Deletion Analysis of GH7 Endoglucanase Gene (cel7B) Promoter Region in a Talaromyces cellulolyticus ligD-Disrupted Strain. Applied biochemistry and biotechnology. 2017 Dec;183(4):1516-1525