Resolving the database sequence discrepancies for the Staphylococcus aureus bacteriophage phi 11 amidase.

Area of Science:

• Microbiology
• Molecular Biology
• Biochemistry

Background:

• Conflicting nucleotide sequences for the Staphylococcus aureus bacteriophage phi 11 amidase gene exist in GenBank.
• Sequence variations and alternative start sites lead to multiple predicted protein sequences.
• Accurate amidase sequence is crucial due to its subcloning, deletion analysis, and experimental use.

Purpose of the Study:

• To resolve the discrepancies in the Staphylococcus aureus bacteriophage phi 11 amidase gene sequences.
• To identify the correct phi 11 amidase protein sequence.
• To support the use of bacteriophage lytic proteins as bactericidal agents.

Main Methods:

• DNA sequence analysis was employed to compare conflicting sequences.
• Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) was used to analyze the purified recombinant phi 11 amidase protein.

Main Results:

• The study identified the correct phi 11 amidase protein sequence.
• Discrepancies in GenBank sequences were resolved through combined analytical methods.

Conclusions:

• The validated phi 11 amidase sequence is essential for accurate research and development.
• Accurate sequence information facilitates the potential application of phi 11 amidase as a bactericidal agent.