The Proteomic Study About Pseudomonas Aeruginosa Biofilm Formation

Purpose:To discover different protein among mucoid PDO300, Pseudomonas aeruginosa wide-type PAO1,PAO1 lasI rhlI mutants and PAO1 lasR rhlR mutants strains.Methods:(1)The same example were prepared to several different protein concentrations.The samples were profiled by SELDI to select best protein concentration of sample.(2)Using CM10 and IMAC3-Cu protein chip to profile the tropina of mucoid PDO300,wide-type PAO1,PAO1 lasI rhlI and PAO1 lasR rhlR mutants strains in order to select a suitable protein chip.(3)The samples were profiled by SELDI then analyzed by Biomarker Wizard software.Results:(1)There were more significant protein peaks when sample concentration was 1.5μg/μl.(2)Analysis of tropina protein fingerprints showed that CM10 chip combined more proteins than IMAC3-Cu chip.(3)Sixteen different proteins were found between mucoid PDO300 and wide-type PAO1(P＜0.01).Ten proteins with M/Z of 4074,3817,8923,8148,3332,4521, 7621,11594,7831 and 13751 were up-regulated while six proteins with M/Z of 6199,6899,12379,6115,6763 and 8576 were down-regulated in the tropina of PDO300.(4)Comparing the tropina of wide-type PAO1 and PAO1 lasI rhlI mutants strain,there was one protein down-regulated in PAO1 lasI rhlI mutants strain(P＜0.05).It's M/Z was 6910.(5)There were four different proteins between wide-type PAO1 and PAO1 lasR rhlR mutants strain(P＜0.05).Two proteins with M/Z of 8924 and 8790 were up-regulated while two proteins with M/Z of 6735 and 6986 were down-regulated in the tropina of PAO1 lasR rhlR mutants strain.Conclusions:(1)The best proteion concentration for SELDI was 1.5μg/μl.(2)CM10 chip could detect more different proteins than IMAC3-Cu and preferably apply to the research of the tropina of Pseudomonas aeruginosa.(3)There were 16 proteins in tropina between mucoid PDO300 and wide-type PAO1.These proteins may contribute to the phenotypic differences between these strains.(4)There was one different protein between wide-type PAO1 and PAO1 lasI rhlI mutants strain.There were four different proteins between wide-type PAO1 and PAO1 lasR rhlR mutants strain.These proteins may have relation to Quorum sensing system.(5)In this study,we have taken advantage of the techniques of SELDI to detect variation in protein expression among different Pseudomonas aeruginosa strains.