E. Mellado et al., A MULTIGENE FAMILY RELATED TO CHITIN SYNTHASE GENES OF YEAST IN THE OPPORTUNISTIC PATHOGEN ASPERGILLUS-FUMIGATUS, MGG. Molecular & general genetics, 246(3), 1995, pp. 353-359
Two approaches were used to isolate fragments of chitin synthase genes
from the opportunistic human pathogen Aspergillus fumigatus. Firstly,
regions of amino acid conservation in chitin synthases of Saccharomyc
es cerevisiae were used to design degenerate primers for amplification
of portions of related genes, and secondly, a segment of the S. cerev
isiae CSD2 gene was used to screen an A. fumigatus lambda genomic DNA
library. The polymerase chain reaction (PCR)-based approach led to the
identification of five different genes, designated chsA, chsB, chsC,
chsD and chsE, chsA, chsB, and chsC fall into Classes I, II and III of
the 'zymogen type' chitin synthases, respectively. The chsD fragment
has approximately 35% amino acid sequence identity to both the zymogen
type genes and the non-zymogen type CSD2 gene, chsF appears to be a h
omologue of CSD2, being 80% identical to CSD2 over 100 amino acids. An
unexpected finding was the isolation by heterologous hybridization of
another gene (chsE), which also has strong sequence similarity (54% i
dentity at the amino acid level over the same region as chsF) to CSD2.
Reverse transcriptase-PCR was used to show that each gene is expresse
d during hyphal growth in submerged cultures.