title: Characteristics of extracellular DNA containing unmethylated CpG motifs isolated from Bacillus subtilis culture medium filtrate.
abstract: The application of CpG DNA is one of the most modern and sufficiently perspective trends in immunotherapy of cancer. The AIM of the investigation was to characterize the DNA as a part of nucleoprotein fraction isolated from Bacillus subtilis culture medium filtrate (CMF) and to evaluate the dynamic of accumulation of such DNA and level of its unmethylated CpG motifs enrichment in the process of cultivation of bacterium.
  
  
  METHODS
  Two bacterial strains--B. subtilis 7025 and B. subtilis GP1-807-03--were used. Polyacrylamide and agarose gel electrophoresis, restriction analysis and transmission electron microscopy were used. The RESULTS of the investigations showed that DNA was accumulated in culture medium irregularly. DNA-containing fractions isolated on the 7th-9th day of cultivation have the greatest sensitivity to restriction endonuclease HpaII. The presence of phages in CMF concentrates has been shown.
  
  
  CONCLUSIONS
  The present findings suggest that 7th-9th day of cultivation are the most optimal to obtain the bacterial extracellular DNA for the following immunotherapy application.
authors.affiliation: R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, National Academy of Sciences of Ukraine, Kyiv 03022, Ukraine. sergeyolishevsky@yahoo.com
authors.forename: Sergiy
authors.initials: S
authors.lastname: Olishevsky
authors.forename: Vyatcheslava
authors.initials: V
authors.lastname: Kozak
authors.forename: Yuriy
authors.initials: Y
authors.lastname: Yanish
authors.forename: Grygoriy
authors.initials: G
authors.lastname: Potebnya
authors.forename: Galyna
authors.initials: G
authors.lastname: Lisovenko
authors.forename: Ivan
authors.initials: I
authors.lastname: Olexienko
authors.forename: Olga
authors.initials: O
authors.lastname: Mazur
authors.forename: Svitlana
authors.initials: S
authors.lastname: Rybalko
authors.forename: Volodymyr
authors.initials: V
authors.lastname: Shlyakhovenko
mesh_terms: D000970:Antineoplastic Agents; D001412:Bacillus subtilis / Q000235:genetics* / Q000276:immunology*; D018899:CpG Islands / Q000235:genetics*; D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis* / Q000302:isolation & purification; D007167:Immunotherapy / Q000639:trends; D013997:Time Factors
chemical_list: D000970:Antineoplastic Agents; D003470:Culture Media; D004269:DNA, Bacterial