شناسايي مولكولي فسفوليپاز B بعنوان يك فاكتور موثر در وير ولانس ميكروارگانيسم

Molecular Detection of Phospholipase B as an Effective Factor in Virulence of Microrganisims

Phospholipases are the major component of all eukariotic bilayer cell membranes and serve as a scaffold for membrane proteins. Phospholipases are a class of ubiquitous enzymes that have in common both their substrate (phospholipid) and the fact that they are all estrase 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 phospholipase B (PLB) search for detection of some fungal PLB was carried out. DNA extraction of Aspergillus fumigatus, A. niger, Fusarium venonatum and Aurobasidium pullulans was carried out and then using degenerate primers based on nocleotide sequences of Phospholipase B were degenerated. Predicted 550 bp product from A. fumigatus was cloned in pGEMT-Easy vector and then transformed into E. coli Top 10 Fʹ competent cell for extraction of cloned DNA fragment. Sequence analysis of 550 bp fragments revealed a sequence for the PLB gene with a high homology to published PLB sequences. Negative result for detection of PLB gene in Fusarium venonatum can be due to unsuitable primer or the lack of PLB gene in the genome of these microrganisim.