ABSTRACT
A new method for selection of bacterium antibiotic resistance genes is proposed and tested for solving the problems related to selection of primers for PCR assay. The method implies clustering of similar nucleotide sequences and selection of group primers for all genes of each cluster. Clustering of resistance genes for six groups of antibiotics (aminoglycosides, ß-lactams, fluoroquinolones, glycopeptides, macrolides and lincosamides, and fusidic acid) was performed. The method was tested for 81 strains of bacteria of different genera isolated from patients (K. pneumoniae, Staphylococcus spp., S. agalactiae, E. faecalis, E. coli, and G. vaginalis). The results obtained by us are comparable to those in the selection of individual genes; this allows reducing the number of primers necessary for maximum coverage of the known antibiotic resistance genes during PCR analysis.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Typing Techniques/methods , DNA Primers/chemical synthesis , DNA, Bacterial/genetics , Drug Resistance, Multiple, Bacterial/genetics , Polymerase Chain Reaction/methods , Aminoglycosides/pharmacology , DNA Primers/genetics , Enterococcus faecalis/drug effects , Enterococcus faecalis/genetics , Enterococcus faecalis/growth & development , Escherichia coli/drug effects , Escherichia coli/genetics , Escherichia coli/growth & development , Fluoroquinolones/pharmacology , Fusidic Acid/pharmacology , Gardnerella vaginalis/drug effects , Gardnerella vaginalis/genetics , Gardnerella vaginalis/growth & development , Glycopeptides/pharmacology , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/growth & development , Lincosamides/pharmacology , Macrolides/pharmacology , Microbial Sensitivity Tests , Multigene Family , Staphylococcus/drug effects , Staphylococcus/genetics , Staphylococcus/growth & development , Streptococcus agalactiae/drug effects , Streptococcus agalactiae/genetics , Streptococcus agalactiae/growth & development , beta-Lactams/pharmacologyABSTRACT
The expression of mRNA of 36 genes involved in implantation was studied by reverse transcription and real-time PCR. Significant differences in mRNA expression during the early and middle stages of the secretion phase were detected for genes mmp7, vegf, il2m, il1ß, il8, il18, tnfα, il10, tgfß, igfbp2, etc.
Subject(s)
Endometrium/metabolism , RNA, Messenger/genetics , Adult , Embryo Implantation/genetics , Female , Humans , Interleukin-10/genetics , Interleukin-1beta/genetics , Matrix Metalloproteinase 7/genetics , Real-Time Polymerase Chain Reaction , Transforming Growth Factor beta/genetics , Tumor Necrosis Factor-alpha/genetics , Vascular Endothelial Growth Factor A/genetics , Young AdultABSTRACT
Certain types of cancer are often correlate with certain chromosomal rearrangements. The chimaeric genes are formed as a result of this rearrangements, and the chimaeric proteins are the products of their expression. The breakpoints of such translocation are often clustered in the genome. Moreover, such breakpoint clusters often contain specific genomic elements like topoisomerase II consensus sites, nuclear matrix attachment regions and DNA sequences, which can make up secondary non-canonical structure. In this review we discuss whether breakpoints may be induced by chromatin structure. Furthermore, we bring up not touched upon literature question about the relation between the breakpoint clusters and the domain organization of corresponding proteins. We also consider possible mechanisms of chromosomal rearrangements.