The objective of this study was to determine the relationship between differential DNA methylation and mRNA expression in uterine leiomyoma by performing a genome-wide analysis

e first methionine of OmpCKP. The OmpCKP is found in the genome of K. pneumoniae KP1_3869, a 1098 bp gene that encodes a polypeptide of 365aa. To define the possible interaction of CpxRKP to the promoter of OmpCKP, we tested whether CpxRKP directly interacts with its promoter region. For that we first cloned and expressed the cpxR gene. The cpxR gene from K. pneumoniae was PCR amplified, cloned into pET-28c and after transformation in E. coli strain BL21, Vorapaxar Expression of the His-tagged protein was monitored following IPTG induction. Cell lysates following purification ” on a Ni-NTA column, where resolved by SDS/PAGE yielded an expected induced band of,24 kDa. Thereafter, gel shift assays were performed using the 32P-labeled ompCKP promoter fragment and CpxR. The protein-DNA complexes after incubation in reaction buffer 16302825” were resolved on 5% PAGE gel and analysis revealed a clear retardation which was proportional to the protein concentration as shown in the CpxAR influences drug efflux to confer antibiotic resistance To decipher whether cpxAR confers antibiotic resistance by affecting drug efflux, screening for a potential efflux phenotype was accomplished by determining the growing ability of NTUHK2044 and NTUH-K2044DcpxAR in the presence of chloramphenicol and CCCP or reserpine as described in methods section. The growth rate of NTUH-K2044DcpxAR in the presence of 0.005 mg/ml chloramphenicol was 2 fold lower than that of NTUH-K2044. Conversely, both wild type and DcpxAR mutant exhibited stunted growth in the presence of chloramphenicol and protonophore CCCP. In independent experiments, growth remained unaltered on the addition of reserpine. Overall, preliminary findings clearly revealed that cpxAR utilises drug efflux as one of the mechanism to confer resistance against antimicrobial compounds such as chloramphenicol. CpxAR confers cross resistance to disinfectants K. pneumoniae is a nosocomial pathogen and has an ability to stay in abiotic surfaces for long; therefore we tested the susceptibilities of NTUH-K2044 and NTUH-K2044DcpxAR towards different concentrations of popularly used disinfectant chlorhexidine and benzalkonium chloride in hospitals. The percent survival of NTUH-K2044DcpxAR was reduced to 50% upon the lowest exposure of chlorhexidine, indicating that cpxAR has a contributory role to mediate disinfectant resistance in this nosocomial pathogen. Outer membrane profile of cpxAR deletion mutant in K. pneumoniae The cell envelope is the prime line for most outside stress conditions that may modify envelope components and thus bring an extra cytoplasmic stress response. In our present study, we found that CpxAR contribute to antibiotic resistance more precisely towards cefepime and chloramphenicol resistance. A reduction in the permeation of antibiotics is generally related to a decrease in porin expression or an alteration in the porin structure. To get an insight, we evaluated the membrane profiles of cpxAR mutant and the wild type. Analysis revealed alterations in both inner and outer membrane fractions of wild type and mutant, however it was intriguing to note the presence of over expressed bands in the outer membrane fractions of cpxAR mutant in varying sizes,30 kDa,,22 kDa and,16 kDa respectively. Expression analysis of the efflux genes in K.pneumoniae Quantitative real-time RT-PCR was used to examine expression of the efflux transporter genes in wild-type, cpxAR mutant, and cpxAR complemented strains. Compared to the wild-ty