Influence Of Ampicillin Resistant Cronobacter Sakazakii On The Stability And Expression Ofgfpuv In Mutant Cells



Construction of Cronobacter sakazakii with pGFPuv could facilitate the tracking of the bacteria in foodstuff. GFPuv plasmids are completed with ampicillin resistant gene for easy selection. Previous studies indicated that ampicillin resistant bacteria could not be made into GFPuv mutants, but the reason was unknown. The objectives of this study were to investigate the performance of C. sakazakii GFPuv mutants based on their stability and GFP expression and to understand the correlation between ampicillin resistance traits in the wild types with the success of GFPuv mutants construction. The results showed that the ampicillin resistance level of C. sakazakii isolates varied. Of the 12 isolates studied, 8 isolates (66.6 %) were susceptible ((FWHc3, FWHb6, FWHb15, YRw3, E2, E4, E6, and E9), two (16.66%) had intermediate resistance to ampicillin (Desb10, E7), and the other two (16.6 %) were ampicillin resistant (FWHd1, E1). The ampicillin sensitive C. sakazakii could be labeled with pGFPuv and their plasmid stability were most stable (E2). The ampicillin resistant C. sakazakii isolates also could be labeled with pGFPuv but the resulting mutants were unstable in the cell and could not express the GFPuv. The high resistance isolate could not express the GFPuv (E7 and FWHd1). Characterization of isolate resistance on antibiotic marker of plasmid is required to assure successfulness and stability of bacteria labelling process.


Cronobacter sakazakii, GFPuv, Mutant,Stability, Ampicillin resistance

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