creators_name: Shaiful Adzni Sharifuddin, creators_name: Sahidan Senafi, creators_name: Nik Marzuki Sidik, creators_id: creators_id: sahidan@pkrisc.cc.ukm.my creators_id: nms@pkrisc.cc.ukm.my type: article datestamp: 2008-07-01 14:15:17 lastmod: 2008-07-01 14:15:17 metadata_visibility: show corp_creators: Universiti Kebangsaan Malaysia. Faculty of Science and Technology. School of Biosciences and Biotechnology corp_creators: Universiti Kebangsaan Malaysia. Faculty of Science and Technology. School of Biosciences and Biotechnology corp_creators: Universiti Kebangsaan Malaysia. Faculty of Science and Technology. School of Biosciences and Biotechnology title: Analysis of DNA Sequence Coding for a Partial Cellobiohydrolase Gene from Aspergillus terreus SUK-1 ispublished: pub subjects: Q subjects: R full_text_status: none keywords: Aspergillus terreus SUK-1, cellobiohydrolase, cellulase, gene, sequence note: This work was supported by IRPA grant 09-02-02-0108-EA260, Ministry of Science, Technology and Innovation, Malaysia abstract: Cellulolytic enzymes are generally composed of multienzyme systems and are divided into three classes, endoglucanase, exoglucanase and β-glucosidase. They work synergistically to hydrolyze cellulose molecules to glucose. Fungal exoglucanase (CBH) is a unique enzyme, capable of degrading highly ordered crystalline cellulose. Here we report the isolation of both genomic and cDNA clone and coding for a partial exoglucanase gene from Aspergillus terreus SUK-1. Two oligonucleotide primers were synthesised based on conserved regions of CBH genes information obtained from database which act as specific primers for gene amplification. PCR amplification was done on both genomic DNA and cDNA. The reactions amplified approximately 538 bp nucleotides from genomic DNA and 486 bp from cDNA which were then sequenced. The sequences were analysed using various platforms such as BLASTX, BLASTN, PSI-BLAST and Pfam. Both genomic and cDNA sequences showed high similarity towards exoglucanase gene and protein from various type of microorganisms. When scanned against Pfam, a high match to glycosyl hydrolase family 7 was observed. Analysis of the genomic sequence showed that it contains a 52 bp intron. date: 2006 date_type: published publication: Malaysian Journal of Biochemistry and Molecular Biology volume: 13 number: 1 publisher: Malaysian Society for Biochemistry and Molecular Biology pagerange: 52-55 refereed: TRUE issn: ISSN 1511-2616 official_url: http://ejum.fsktm.um.edu.my/ArticleInformation.aspx?ArticleID=594 referencetext: 1. Knowles, J., Lehtovaara, P. and Teeri, T. Cellulase families and their genes. Trends Biotech 1987; 5: 255-260. 2. Henrissat, B. Glycosidase families. Biochem J Biochem Soc Trans 1998; 26:153-156. 3. Kader, J., Othman, O. and Zainal, A. Kajian ke atas kulat selulolisis Aspergillus terreus SUK-1. Bangi: Imperatif dalam penyelidikan sains hayat. 1988; p. 221-224 4. Othman, O., Jalil A.K. and Al-Arikah, R.A. Studies on extracellular cellulose during growth of Aspergillus terreus SUK-1. Sains Malay 1984; 13: 341-354. 5. Mandel, M. and Reese, E.T. Induction of cellulose in fungi by cellobiose. J Bacteriol 1957; 73: 816-826. 6. Sambrook, J., Fritsch, E. and Maniatis, T. Molecular cloning: A laboratory manual, 2nd ed. 1989. Cold Spring Harbor Laboratory, New York. 7. Pretea, M., Lin, X. and Salzberg, S.L. GeneSplicer: a new computational method for splice site prediction. Nucleic Acids Res 2001; 29: 1185-1190. 8. Kitamura-Abe, S., Takahashi, N., Itoh, H., Sakurai, A., Saito, R., Washio, T., Shimbo,M., Tsutsumi, A. and Tomita, M. Distribution analysis of the oligonucleotide around 5‘ splice sites in H. sapiens, M. musculus, D. Melanogaster and A. thaliana pre-mRNAs. Gen Inform 2002; 13: 428-429. 9. Keller, E.B. and Noon W.A. Intron splicing: a conserved internal signal in introns of Drosophila pre-mRNAs. Nucleic Acids Res 1985; 13: 4971-4981. 10. Sims, P., James, C. and Broda, P. The identification, molecular cloning and characterization of a gene from Phanerochaete chrysosporium that shows strong homology to the exo-cellobiohydrolase I gene from Trichoderma reesei. Gene 1988; 74: 411-412. 11. Gielkens, M.M., Dekkers, E., Visser, J. and de Graaff, L.H. Two cellobiohydrolase-encoding genes from Aspergillus niger require D-xylose and the xylanolytic transcriptional activator XlnR for their expression. Appl Environ Microbiol 1999; 65: 4340-4345. 12. Hong, J., Tamaki, H., Yamamoto, K. and Kumagai, H. Cloning of a gene encoding thermostable cellobiohydrolase from Thermoascus aurantiacus and its expression in yeast. Appl Microbiol Biotechnol 2003; 63: 42-50. 13. Takada, G., Kawaguchi, T., Sumitami, J. and Arai, M. Cloning, nucleotide sequence, and transcriptional analysis of Aspergillus aculeatus No. F-50 cellobiohydrolase (cbhI) gene. J Fermen Bioeng 1998; 85: 1-9. 14. Lockington, R.A., Rodbourn, L., Barnett, S., Carter, C.J. and Kelly, J.M. Regulation by carbon and nitrogen sources of a family of cellulases in Aspergillus nidulans. Fungal Genet Biol 2002; 37: 190-196. citation: Shaiful Adzni Sharifuddin, and Sahidan Senafi, and Nik Marzuki Sidik, (2006) Analysis of DNA Sequence Coding for a Partial Cellobiohydrolase Gene from Aspergillus terreus SUK-1. Malaysian Journal of Biochemistry and Molecular Biology, 13 (1). pp. 52-55. ISSN ISSN 1511-2616