Cloning and Sequencing of sacol a novel gene from Staphylococcus aureus S Menbari, MR Pourmand, MH Shirazi, N Mardani IJMM 2008; 2(1):9-14 ICID: 873185
Article type: Original article
IC™ Value: 3.25
Abstract provided by Publisher
Background & Objectives: Natural staphylococcal infections and vaccines based whole bacteria lead to poor antibody responses, but recent research reveals that specific antibodies based on recombinant staphylococcal antigens are much more protective. Sacol is a novel antigen that its structural and immunological traits poorly characterized. This research aimed to clone of sacol, a novel gene from Staphylococcus aureus.
Material and Methods: The specific primers with suitable restriction sites were designed and sacol amplified by PCR. The sacol and plasmid were produced as sticky ends by restriction enzymes NdeI and XhoI. To amplify the recombinant plasmid the pET21sacol transferred into competent cell E.coliTOP10. The recombinant plasmid harvested from the host and analyzed by restriction enzymes and sequencing. Finally, sacol gene analyzed by bioinformatics tools.
Results: The sacol gene has 723bp which amplified, cloned and sequenced successfully. Sacol is highly conserved in Staphylococcus aureus strains. Moreover, software analysis shows that sacol encodes a protein with 32KDa molecular weight (267 amino acids) which has similarity with C51 peptidase in N-terminal with one alpha helix and 14 beta sheets.
Conclusion: the sacol gene is conserved in majority of Staphylococcus aureus strains and may exist and express in most of staphylococcal infections.The role and regulation of the gene is thus of great interest.
Keywords: Staphylococcus aureus, sacol, cloning