Abstract: Molecular cloning and functional research on genes related to fungal taxol biosynthesis is a key for the genetic engineering improvement of taxol-producing fungi. In this study, the full length sequences (1674 bp) of DNA and cDNA of Unigene A09801 were cloned from the taxol-producing fungus, Cladosporium cladosporioides MD2, respectively. Unigene A09801 sequence had no introns. Unigene A09801 encoding protein contained 557 amino acids, had 56% identity with GMC (glucose-methanol-choline) oxidoreductase, possessed 61 291.8 u molecular weight and 5.47 isoelectric point value, and had no trans-membrane structure and signal peptide. Unigene A09801 existed in the fungal genome with a single copy. The expression levels of Unigene A09801 responded to Methyl Jasmonate were showed to decrease firstly, increase subsequently and then decrease, and the highest expression level appeared at 24 h. The prokaryotic expression vector of Unigene A09801 was constructed, and the optimized expression conditions of Unigene A09801 in E. coli BL 21 were: induced at 32℃ for 4 h, with 1.2 mmol/L TPTG. These results would lay a foundation for further in vitro analyzing catalytic function of Unigene A09801 encoding protein.

Key words: Cladosporium cladosporioides, taxol, synthase, prokaryotic expression