A silent antifungal protein (AFP)-like gene lacking two introns in the mould Trichoderma viride

In cells of the mould Trichoderma viride, the existence of an antifungal pr otein (AFP)-like gene consisting of 285 bp was confirmed by Southern analys is that genomic DNA of T. viride could hybridize with the cDNA of mature AF P of Aspergillus giganteus MDH 18894. Except for the absence of two introns , the nucleotide sequence of the AFP-like gene was identical to that of the AFP gene of A. giganteus in positions 336-479, 568-649, and 706-765. The A FP-like gene could not be transcribed into its mRNA in T. viride cells as e xamined by RT-CR using total RNAs of T. viride as template. Furthermore, AF P could not be detected either directly from the culture medium of T. virid e or by Western analysis. However, the AFP-like gene could be actively expr essed like the cDNA of AFP in Escherichia coli cell. Recombinant AFP exhibi ted similar antifungal activity as native AFP. (C) 2000 Elsevier Science B. V. All rights reserved.