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Objective: research siHybrids technology against Pseudomonas aeruginosa the PAO1 the efflux pump mexB gene silencing effect in vitro . Method : 1. Against Pseudomonas aeruginosa strain PAO1 wild efflux pump mexB gene design and synthesis of specific siHybrids siHybrids molecules and a negative control molecule . At the molecular concentration 50nM , the interference synthesis design siHybrids molecules Pseudomonas aeruginosa PAO1 , and set up the experimental group Pseudomonas aeruginosa PAO1 blank control group, negative control group scambl ( ESC ) -001 intervention group siHybrids (si) -001, siHybrids (si) -002 and -003 siHybrids ( si ) 3 , respectively, in the intervention after 12 hours and 24 hours after each experimental group of Chinese and foreign real - time quantitative PCR the efflux pump target genes MexB [ 1 ] mRNA expression levels. 4 siHybrids molecules using Mueller-Hinton fold dilution assay concentration of 50nM against Pseudomonas aeruginosa PAO1 and chloramphenicol ( CP ) , erythromycin ( EM ) , before and after the action of levofloxacin ( L - OFLX ) intervention MIC ( minimum inhibitory concentration ) . results: 1. different siHybrids molecules intervention in the PAO1 12 hours after , MexB gene mRNA expression was no significant difference . Intervention after 24 hours (si-001, si-002, si-003) the MexB gene mRNA expression intervention group than the control group , negative control group ( sc - 001 ) has decreased significantly. Of comparison intervention 12 hours , 24 hours after mexB gene mRNA expression blank control group, negative control group ( sc - 001 ) , you can find the amount of mRNA expression tended to increase , while the intervention group ( si - 001 , si - 002 , SI - 003 ) the MexB gene mRNA expression decreased. Molecules 4.siHybrids intervention before and after 24 hours of chloramphenicol ( CP ) , erythromycin ( EM ) , the MIC of levofloxacin ( L - OFLX ) no significant difference Conclusion : mRNA expression levels , siHybrids molecular intervention Pseudomonas in vitro Pseudomonas the PAO1 mexB mRNA expression , such silencing effect showed a time-dependent manner , can effectively play the role of intervention within 24 hours .
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