Identification of Phospholipase D Gene as a Contributing Factor in Growth and Virulence of Microorganisms

Secretory extracellular Phospholipases are generally involved in hydrolysis of extracellular phospholipids and thus providing nutritive source of carbon, nitrogen, and phosphate. However, intracellular phospholipases perform metabolic functions and adjust biologic activities. Synthesis of phospholipases in different pathogenic microorganisms and their mode of action in virulence of the microorganism have been the center of attention in recent studies. During this study using degenerate primers based on homologous amino acid sequences of phospholipase D (PLD) search for detection of Aspergillus fumigatus was carried out. DNA extraction of A. fumigatus was performed and then using degenerate primers based on nucleotide sequences of Phospholipase D gene was degenerated. Predicted 850 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 850 bp fragments revealed a sequence for the PLD gene of A. fumigatus with a high homology to published PLD sequences in other microorganisms. Gene sequence studies are generally conducted to determine the participation of gene expression in pathogenecity of microorganisms, the evaluation of biochemical features and physiologic function of gene product for understanding the basic knowledge to provide immunity, production of vaccine, drug, or blocker for gene product, utilizing the gene product, utilizing the gene product for infection detection and so on. Thus, cloning of a part of phospholipase D gene in order to full identification of nucleotides sequence of this gene could contribute to achieve those goals.