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

• Published in: Biochimica et biophysica acta Vol. 1475; no. 2; pp. 119 - 124
• Main Authors: Hao, Jian-Jiang, Ye, Jun-qiang, Yang, Qiang, Gong, Zhen-zhen, Liu, Wang-Yi, Wang, En-duo
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 03.07.2000
• Subjects: AFP-like gene; Amino Acid Sequence; analysis; Aspergillus; Aspergillus - genetics; Base Sequence; biosynthesis; DNA, Complementary; DNA, Complementary - biosynthesis; DNA, Complementary - isolation & purification; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Escherichia coli - genetics; Fungal Proteins; Fungal Proteins - analysis; Fungal Proteins - genetics; Gene Silencing; Genes, Fungal; Genetic Vectors; genetics; Intronless; Introns; isolation & purification; Molecular Sequence Data; Recombinant Proteins; Recombinant Proteins - analysis; Reverse Transcriptase Polymerase Chain Reaction; Sequence Alignment; Transcription, Genetic; Trichoderma; Trichoderma - genetics; Trichoderma viride; Intronless; AFP-like gene; Trichoderma viride
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/S0304-4165(00)00047-7

In cells of the mould Trichoderma viride, the existence of an antifungal protein (AFP)-like gene consisting of 285 bp was confirmed by Southern analysis that genomic DNA of T. viride could hybridize with the cDNA of mature AFP 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 AFP-like gene could not be transcribed into its mRNA in T. viride cells as examined by RT-PCR using total RNAs of T. viride as template. Furthermore, AFP could not be detected either directly from the culture medium of T. viride or by Western analysis. However, the AFP-like gene could be actively expressed like the cDNA of AFP in Escherichia coli cell. Recombinant AFP exhibited similar antifungal activity as native AFP.