In vitro antibacterial activity of recombinant protein AtlM of Staphylococcal aureus
2013
Objective To construct the recombinant protein AtlM (rAtlM) of Staphylococcal au- reus through prokaryotic expression system and to investigate its antibacterial activity in vitro. Methods The specific primers were designed according to atlM gene sequence of Staphylococcal aureus recorded in Gen- Bank, and atlM gene was amplified by PCR from the Staphylococcal aureus strain (ATCC25923). The re- combinant plasmid pET-32a (+)/atIM was constructed and transformed into Transetta (DE3) to express AtlM after induced by IPTG. The expressed protein AtlM was further analyzed by SDS-PAGE and purified by electroeluting of bag filter. The minimal inhibitory concentrations (MICs) of rAtlM to ATCC25923 and oxac- illin-resistant S. aureus strain were determined by the broth microdilution method. S. aureus ATCC25923 strain and oxacillin-resistant S. aureus strain (final concentration of 5×10^5 CFU/ml) were exposed to rAtlM (50 μg/ml) respectively to test its antibacterial activity in vitro. Results The recombinant protein AtlM was expressed and purified successfully with a relative molecular weight of 80×10^3 and a concentration of 1.25 mg/L. The MICs of rAtlM to ATCC25923 strain and oxacillin-resistant S. aureus strain were 8 μg/ml and 64μg/ml, respectively. In vitro test showed that rAtlM had inhibitory effects on the growth of ATCC25923 strain and oxacillin-resistant S. aureus strain after 1 h of intervention (P=0.004 and P=0.026, respectively), which lasted to 5 h for ATCC25923 strain (P = 0. 012) and 3 h for oxacillin-resistant strain (P= 0. 001 ). Conclusion This study shows that rAtlM has a certain antibacterial effects on S. aureus ATCC25923 strain and oxacillin-resistant S. aureus strain, suggesting a possibility of serving as antimicrobial agent.
Key words:
Staphylococcal aureus ; Autolysin ; Antimicrobial agent; Gene expression
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