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B P Ulanov

Publications and source records attributed to B P Ulanov.

At least 19 recordsLinked to original sources

[Z-form of intraphage DNA].

The bacteriophage lambda gt10 DNA containing an insertion of 20 pairs of GC-bases capable of being arranged as Z-form was cloned. Two independent methodological approaches based on the main properties of Z-form were used to study the shape of the insertion: formation of transition bridges composed of unpaired nucleotides between left-rotating Z-forms and right-rotating B-forms of helix (j-domain) and high immunogenic activity of Z-form. O-beta-diethylaminoethylhydroxyamine (OHA), an analogue of hydroxylamine, is capable of reacting specifically with unpaired cytidines. In this work this modification was used to inhibit the process of restriction at BamH1-site adjacent to (gc)10 insertion, that N-Methyl-bis(2-chloethyl) amine (MBCA) is capable of fixing the Z-form of the insertion in situ. Fixed Z-form is conserved even after DNA has been isolated from bacteriophage, thereby providing an opportunity of its identification by anti-Z-antibodies. It was shown that from 4 to 6% of the total number of insertions are in the Z-form. The hypothesis of significant role of Z-form in the process of packing of DNA molecules in capsid is put forward.

Bacteriophage lambda↗

[The Z-form of bacteriophage lambda DNA, modified in situ].

Using the methods of chemical modification, restriction analysis and immune-electron microscopy it has been shown that the definite regions of the bacteriophage lambda DNA contain unpaired bases in situ. The distribution map of such sites along the genome has been constructed. The correlation of the in situ modification and the reaction with anti-Z-DNA antibodies is shown for the 44972 bp site of bacteriophage DNA. The possibility of the existence of Z-form DNA in situ is discussed.

Bacteriophage lambda↗

[Unpaired bases in the B--Z junction of the supercoiled plasmid].

The restriction analysis has been used to establish that O-beta-diethylaminoethylhydroxylamine (OHA) produces modification of unpaired cytidines in the polylinker region adjacent to the Z-insert (dG-dC)10. (dG-dC)10 in the negatively supercoiled plasmid pGC20. The length of the transition region between B- and Z-portions of DNA is not less than 36 bps. The reaction of OHA with the unpaired cytidines in the B-Z junction is a fixing one and produces no secondary despiralling of the neighboring regions. The reaction with DNA proceeds much slower than the one with monomers and single-strand polynucleotides. The structural nonuniformity has been observed, which is manifested in the alternating B and "non-B" form DNA in the B-Z junction. It is suggested that these junctions may contain nucleotide sequences which are stable to violation of the B structure during the change in superhelical density of DNA.

Base Sequence↗

[Properties of the plasmid pFT15/10-1 isolated from the vaccine strain of Francisella tularensis].

Plasmid, designated pFT15/10-1, was isolated from Francisella tularensis vaccine strain 15/10. The plasmid is presented by the homogeneous 5.02 +/- 0.054 Md monomeric circular DNA molecules in electron microscopic preparations. Plasmid size is 7-7.3 kb as defined by electrophoresis in agarose gel. The restriction analysis has revealed that plasmid pFT15/10-1 possesses a single specific cleavage site for restriction endonuclease EcoRI, two sites for restriction endonucleases BamHI, BgIII, HincII, HindIII, PstI, three sites for BglI and SalI, some for AluI, TagI, MvaI, CfrI. Plasmid is not digested by restriction endonucleases SmaI, XmaI, KpnI, MluI. Restriction map of the plasmid was constructed for most frequently used restriction endonucleases.

Bacterial Vaccines↗

[Double modification of cytidine residues in DNA].

The reaction of O-beta-diethylaminoethylhydroxylamine (O-beta-HA) with cytidine was studied and its mechanism was shown to be analogous to that of the reaction of hydroxylamine of O-methylhydroxylamine with cytidine. In experiments involving reaction of denatured DNA with O-beta-HA., Sephadex G-15 columns were used for the quantitative separation of normal and modified nucleosides after enzymatic hydrolysis of modified DNA by exonuclease A5 followed by alkaline phosphatase treatment. DNA cytidine residues of free cytidine with O-beta-HA. Modified cytidines can form complex with phosphotungstic acid (PTA). It was shown that one mole of PTA was bound per one mole of modified cytidine either in DNA or in free state. Electron microscopic examination of denatured DNA molecules modified by O-beta-HA and reacted with PTA revealed linear arrays of electron-scattering spots which presumably correspond to PTA molecules complexed with modified cytidine in DNA chains.

Alkaline Phosphatase↗