| |  | | | | | | BlsI- and GlaI-PCR assays – a new method of DNA methylation study | |
| | BlsI- and GlaI-PCR assays have been developed to study DNA methylation. A new method includes DNA hydrolysis by methyl-directed site-specific DNA endonucleases GlaI or BlsI with subsequent routine PCR. Study of DNA methylation in regulation region of human tumor suppressor genes has been performed for a new method evaluation. BlsI- and GlaI-PCR assays have revealed different methylation patterns in promoter region of DAPK1, in promoter and first exon region of RARB and in first exon region of RASSF1A tumor suppressor genes in malignant cell lines HeLa, Raji, U-937, Jurkat and control L-68 cells. GlaI-PCR assay has shown methylation of RARB promoter and first exon region in DNA from all malignant cell lines, but not in control L-68 cells. GlaI- and BlsI-PCR assays have displayed DNA methylation of RASSF1A first exon region in Raji and Jurkat cells only. BlsI-PCR assay of DAPK1 promoter region has demonstrated an additional DNA methylation in Raji cells only. GlaI- and BlsI-PCR assays may be useful both in determination of malignant cells and their discrimination. | |
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| | | | | The resctriction endonucleases detection in colonies of microorganisms Streptomyces and Nocardia | |
| | A simple technique is proposed for detection of restriction endonucleases in bacteria Streptomyces and Nocardia. The analysis was performed directly in the cells cultivated on Petri dishes and collected from colonies by an inoculation loop. The cells were treated with lysozyme, EDTA and Triton X-100. The lysates were tested for restriction endonucleases. The technique enables todetect the enzymes Nco I, Not I, Nru I I, Sfr 3031, and Sfi I in the lysates of corresponding strains-producers. | |
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| | | | | A simple technique for detection of restriction endonucleases in bacterial colonies | |
| | A simple technique is proposed for detection of bacterial restriction endonucleases. Analysis is performed directly in the cells from colonies cultivated on Petri dishes. The cells are collected by a microbiological loop into microtubes and then are treated with lysozyme and Triton X-100. After centrifugation of bacterial lysat the supernatant is tested for a presence of site-specific endonuclease activity. The technique enables to analyze about 100 colonies for 3-4 hours. | |
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