Submitted to: Journal of Basic Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/18/2007
Publication Date: 2/1/2008
Publication URL: http://www3.interscience.wiley.com/cgi-bin/fulltext/117905904/PDFSTART
Citation: Donovan D.M., Foster-Frey J., Garrett W.M., Blomberg, L. 2008. Resolving the database sequence discrepancies for the Staphylococcus aureus bacteriophage phi11 amidase. Journal of Basic Microbiology. 48(1):48-52. Interpretive Summary: Bacteriophage endolysins are proteins that digest bacterial cell walls and thus are a new source of antimicrobials. Work from several labs indicates that the phage phi11 endolysin is a potent antimicrobial that can kill Staphylococcus aureus and several coagulase negative staphylococcal pathogens. However, there are multiple conflicting Genbank submissions of the DNA (and predicted protein sequence) of this gene. Also, multiple versions have been subcloned and used in a recombinant form from different labs. We have performed DNA sequence analysis and MALDI-TOF mass spec. analysis on the purified protein, to verify the correct DNA and protein sequence of this gene and its product.
Technical Abstract: There are two conflicting primary nucleotide sequences of the Staphylococcus aureus bacteriophage '11 amidase gene in public databases. Nucleotide sequence differences as well as alternative translational start site assignments result in three non-identical protein sequence predictions in Genbank for this amidase. Multiple versions of the amidase gene have been subcloned, deletion analysis performed, and their experimental use described. Due to a resurgence of interest in the expression and use of bacteriophage proteins as bactericidal agents and the high antimicrobial potential of the phi11 amidase, it is prudent to verify the correct phi11 amidase protein sequence. The protein sequence discrepancies are resolved through a combination of DNA sequence analysis and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry analysis of the recombinant purified phi11 amidase protein.