Abstract
Endo-β-glucanase II (EG II) gene cDNA was isolated from the fungus Humicola insolens H31-3 by RT-PCR. It was cloned into the expression vector pGAPZαA. The resultant recombinant plasmid was introduced into Pichia pastoris GS115 by electroporation after being linearized by BspHI digestion. The recombinant Pichia pastoris strain was obtained and SDS-PAGE showed that the molecular weight of the expression protein was about 55 kD.The cultivation condition and the characteristics of the recombinant EG II were also explored.
Similar content being viewed by others
References
Dalboge H, Heldt-Hansen H P (1994). A novel method for efficient expression cloning of fungal enzyme genes, Mol Gen Genet, 243: 253–260
Ghose T K (1987). Measurement of cellulase activity, Pure and Appl Chem, 59(2):257–268
Han S J, Yoo Y J, Kang H S (1995). Characterization of a bifunctional cellulase and Its structural gene, J Biol Chem, 270(43): 26012–26019
Moriya T, Watanabe M, Sumida N, Okakur A, Murakami T (2003). Cloning and overexpression of the avi2 gene encoding a major cellulase produced by Humicola insolens, Biosci Biotech Biochem, 67(6): 1434–1437
Sambrook J, Russell DW (2002). Molecular Cloning: A Laboratory Manual, 3rd ed., New York: Cold Spring Harbor Laboratory Press, 485–487
Schulein M (1997). Enzymatic properties of cellulases from Humicola insolens, J Biotech, 57: 71–81
Takashima S, Nakamura A, Masaki H, Hidaka M, Uozumi T (1996). Cloning, sequencing, and expression of the cellulase genes of Humicola grisea var. thermoidea, J Biotech, 50: 137–147
Takashima S, Nakamura A, Masaki H, Uozumi T (1996). Purification and characterization of cellulases from Humicola grisea, Biosci Biotech Biochem, 60(1): 77–82
Takashima S, Nakamura A, Masaki H, Uozumi T (1997). Cloning, sequencing, and expression of a thermostable cellulase gene of humicola grisea, Biosci Biotech Biochem, 61(2): 245–250
Wang J (1997). Study on technological parameters of washing jean using cellulase, Journal of Anhui Agricultural University, 24(1): 84–87 (in Chinese)
Wang J Z, Fang M (2000). The Protein Protocols Handbook. Beijing: Science Press (in Chinese)
Waterham H R, Digan M E, Koutz P J, Lair S V, Cregg J M (1997). Isolation of the Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase gene and regulation and use of its prometer, Gene, 186: 37–44
Xiao C L, Xu C X (2002). Application study on microbial cellulase, Journal of Microbiology, 22(2): 33–35 (in Chinese)
Author information
Authors and Affiliations
Corresponding author
Additional information
__________
Translated from Microbiology, 2006, 33(6): 68273 [译自: 微生物学 通报]
Rights and permissions
About this article
Cite this article
Tan, H., Zhang, G., Zheng, G. et al. Cloning and expression of endo-β-glucanase II gene in Humicola insolens H31-3. Front. Biol. China 3, 287–292 (2008). https://doi.org/10.1007/s11515-008-0053-7
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11515-008-0053-7