MOLECULAR CLONING OF THE GENE CODING FOR CELLOBIOHYDRASE II FROM TRICHODERMA VIRIDE
绿色木霉纤维素酶CBHII基因的分子克隆

Gene library of Trichoderma viride was constructed in lambda EMBL3 DNA vector and screened by three rounds of plaque in situ hybridization with two probes representing 5'- or 3'-end sequences of cbh2 gene of Trichoderma reesei. Three of five positive clones were further tested by cross dot blotting with four probes respectively from 5'- or 3'-end sequences of cbh1 and cbh2 genes of Trichoderma reesei. It was confirmed that full-length cbh2 gene of Trichoderma viride had been cloned. However, homology in the end sequences had not been found between cbh1 and cbh2 genes of the Trichoderma. The length of inserts in vector EMBL3 DNA arms was identified by restriction enzyme analysis, and one of the inserts was then cloned into plasmid pUC19. It was convinced by southern blotting that the resultant recombinant plasmid (pCBH II-14) had contained cbh2 gene of T. viride.