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High Throughput Screening of Fungal Endoglucanase Activity in Escherichia coli
Published on: August 13, 2011
Isolation and characterization of the Aspergillus nidulans eglC gene encoding a putative beta-1,3-endoglucanase
Chang-Jun Choi1, Hee-Jeong Ju, Byung-Hyun Park
1Division of Biological Sciences, Basic Science Research Institute, Chonbuk National University, Chonju, Chonbuk 561-756, Republic of Korea.
Abstract:
The Aspergillus nidulans eglC gene, which encodes a putative beta-1,3-endoglucanase, was isolated from a chromosome-specific library by using an expressed sequence tag, esd0113. The EglC open reading frame encodes a 465 amino acid polypeptide, of which the amino acid sequence showed 46% similarity to that of Saccharomyces cerevisiae beta-1,3-endoglucanase. The eglC transcript level at the early stages of asexual and sexual developments was dependent on the presence of the nsdD gene that encodes a GATA-type transcription factor, confirming that the nsdD gene is necessary for full accumulation of the eglC transcript. Deletion of the eglC gene did not affect the radial growth rate, the germination rate of conidia, and both of asexual and sexual development. However, deletion of the gene led to hyphae more resistant to a cell wall-lyzing enzyme, implying that the cell wall structure of the eglC-null mutant is altered from a wild type one. Furthermore, deletion of the fadA and sfaD genes, that encode a Galpha and a Gbeta subunits of a heterotrimeric G protein, respectively, did not affect the eglC transcript level at the early developmental stages. In contrast, deletion of the flbA gene, that codes for a regulatory protein having an RGS (regulator of G protein signaling) motif, led to decrease in the eglC transcript level. The eglC transcript level was not higher in a creA mutant than in a wild type, indicating that the eglC gene is not sensitive to carbon-catabolite repression.
