Abstract：The cellobiohydrolase gene cbh1 fragment (GenBank Accession No. AY840982) was amplified by RT-PCR from thermophilic fungus Thermoascus aurantiacus var. levisporus. RACE was used to obtain its full-length cDNA (1710 bp), encoding 457 amino acids. The first 19 amino acids of the deduced amino acid sequence were presumed to be a signal peptide. The fragment encoding mature cellobiohydrolase was inserted into Pichia pastoris vector pPIC9K to construct recombinant plasmid pPIC9K/cbh1. The pPIC9K/cbh1 was then introduced into Pichia pastoris GS115 and 61 transformants were obtained, After comfirmed by G418 risistance and PCR, and induced expression, one clone GSp-15 was selected from the 61 transformants. The expression level of GSp-15 was 1.17 mg/mL after induction for 144 h in methanol, and its activity was 20.3 U/mL with p-NPC as substrate.
陈静;李多川;张玉芹;杜欣可. 嗜热子囊菌光孢变种cbh1基因的cDNA克隆及在毕赤酵母的高效表达[J]. , 2006, 14(3): 406-411.
CHEN Jing;LI Duo-chuan;ZHANG Yu-qin;DU Xin-ke. cDNA Cloning of the Cellobiohydrolase Gene cbh1 from hermoascus aurantiacus var. levisporus and Its Expression in Pichia pastoris . , 2006, 14(3): 406-411.
