Screening for anti-multidrug associated genes of Shigella flexneri with suppression subtractive hybridization

ObjectiveTo investigate the differential genes between the induced anti-multidrug strain and sensitive strain of Shigella flexneri with suppression subtractive hybridization(SSH) and to screen the genes related to multidrug resistance and to analyze its relationship with induced drug-resistance.MethodsThe induced anti-multidrug subtractive DNA library was constructed from the DNA of anti-multidrug strain(tester) and sensitive strain of Shigella flexneri(driver).The parts of positive clones in the library were identified by PCR,screened by dot blotting hybridization,and then sequenced and analyzed with the BLAST software in GenBank.ResultsThe induced anti-multidrug subtractive DNA library was constructed successfully.A total of 12 induced multidrug resistant differential fragments were acquired,in which,6 fragments were sequenced and compared with the GenBank Database,3 fragments failed to match with the homologous genes in Genbank,while the other 3 fragments had a high similarity to known genes(16S rRNA gene,hypothetical Rep protein,and type I site-specific deoxyribonuclease).ConclusionAnti-multidrug of Shigella flexneri is a multi-genes involved in a complex process and the screened novel gene fragments could provide basis for further cloning of full-length genes.