17-23: Analysis of gene expression in Acremonium cellulolyticus using High-coverage gene expression profiling (HiCEP) method

Tuesday, May 1, 2012
Napoleon Ballroom C-D, 3rd fl (Sheraton New Orleans)
Akihiro Hideno, Senior Research Fellow Center, Ehime University, Matsuyama, Japan, Hiroyuki Inoue, Biomass Technology Research Center, National Institute of Advanced Industrial Science and Technology, Higashi-hiroshima, Japan, Tatsuya Fujii, Biomass Technology Research Center, National Institute of Advanced Industrial Science and Technology, Higashi-Hiroshima, Japan, Shinichi Yano, Biomass Technology Research Center, National Institute of Advanced Industrial Science and Technology, Hiroshima, Japan and Shigeki Sawayama, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
The development of hyper cellulases producing filamentous fungi has been performed all over the world because the reduction of cost for cellulase enzymes has been required in ethanol production from lignocellulosic biomass. Acremonium cellulolyticus, which was isolated from soil in Japan, secretes not only a lot of cellulase but also β-glucosidase. This strain had been bred by mutagenesis with UV and nitrosoguanidine (NTG) during more than 20 years, and the CF-2612 strain has been developed recently. To improve the capability of cellulase production further of this strain, the introduction of genetic engineering is essential. However, there is very little genetic information of A. cellulolyticus, and the target genes for breeding can be limited. In this study, we present a global analysis of specifically-expressed genes with cellulose using HiCEP method to obtain the genetic information for cellulase production in CF-2612 of the most productive strain. The CF-2612 strain was cultivated for 16-18 h using three kinds of carbon source; glycerol (Gly), lactose (Lac), and Solka Floc (SF). The total RNA was extracted by this growth cell, and the comparison of expression was performed by the method of HiCEP. Approximately 15,000 peaks were obtained from using each carbon source culture, respectively. Thirty four genes were selected and sequenced based on the intensities as the expression levels of cultures with; Gly <SF and Lac <SF and were annotated. The expression levels of 34 genes were confirmed by the method of quantitative real-time PCR.
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