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Several transformants was isolated and cultivated to confirm the production of rAaBGL1.

Plasmid DNA from positive transformants was isolated and sequenced.

The genomic DNA of G418-resistant transformants was isolated for verifying the integration by PCR using pairs of primers (Table 1): Sp-N and Sp-C, up-N and down-C, kc-1 and kc-2, and kc-3 and kc-4, respectively.

Genomic DNA from the transformants was isolated as described earlier [ 20].

Yeast chromosomal DNA of transformants was isolated and PCR controls were performed.

Yeast chromosomal DNA of transformants was isolated and three PCR controls were performed.

Yeast were transformed using lithium acetate, and transformants were isolated after growth on solid selective medium (lacking histidine).

Transformants were isolated, and the transformed kanamycin resistance gene (kan) was detected from the plasmid DNA extracts of the recombinant strains by PCR and was confirmed by restriction digestions.

The wild-type strain Aa113 was transformed with pBSKC1, and 26 HR transformants were isolated.

The resultant construct was transformed into E. coli BL21 (DE3) and transformants were isolated at 37°C on Luria-Bertani (LB) agarose media containing 100 μg/mL ampicillin.

After transformation of the Δ xyr1 strain, several independent transformants were isolated based on their recovered ability to form hydrolytic halos on Avicel plate (data not shown).