[Molecular detection of phospholipase B3 as an effective factor in virulence of Aspergillus fumigatus]

ABSTRACT

Phospholipases are the major component of all eukaryotic bilayer cell membranes. and serve as a scaffold for membranes proteins. Phospholipases are a class of ubiquitous enzymes that have estrase activity and due to their ability to lyses of cell membrane can play an important role for pathogenicity of microrganisims. During this study using degenerate primers based on homologous amino acid sequences of some fungi phospholipase B [PLB], detection of Aspergillus fumigatus PLB3 was carried out. After DNA extraction of A. fumigatus, A. niger, Fusarium venonatum and Aurobasidium pullulans PCR reaction was carried out using degenerate primers. Predicted 2100 bp product from A. fumigatus was cloned in pGEMT-Easy vector and then transformed into E. coli Top 10 F' competent cell for extraction of cloned DNA fragment. Sequence analysis of 2100 bp fragments using BLASTX software were revealed a high homology to published PLB sequences from other fungi. Phylogenic tree analysis of PLB3 gene shows that PLB and potential PLB analogues like lysophospholipase are contained in a large cluster of the PLB family. A. fumigatus PLB gene is more closely related to A. oryzae, A. niger, Penicillium chrysogenum and Neurospora crassa than other fungi. Also, sequence availability of full length PLB3 gene represents a noteworthy breakthrough in the study of this opportunistic pathogen and function of PLB3 gene in pathogenesis of A. fumigates. Detection of PLB3 gene can be lead to preparing vaccine, blocker and laboratory marker for recognition of PLB3 gene products either in patient body or clinical samples