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O Benada

Publications and source records attributed to O Benada.

26 records · Page 2Linked to original sources

Characterization of new plasmids from methylotrophic bacteria.

Several tens of methanol-utilizing bacterial strains isolated from soil were screened for the presence of plasmids. From the obligate methylotroph Methylomonas sp. strain R103a plasmid pIH36 (36 kb) was isolated and its restriction map was constructed. In pink-pigmented facultative methylotrophs (PPFM), belonging to the genus Methylobacterium four plasmids were detected: plasmids pIB200 (200 kb) and pIB14 (14 kb) in the strain R15d and plasmids pWU14 (14 kb) and pWU7 (7.8 kb) in the strain M17. Because of the small size and the presence of several unique REN sites (HindIII, EcoRI, NcoI), plasmid pWU7 was chosen for the construction of a vector for cloning in methylotrophs. Cointegrates pKWU7A and pKWU7B were formed between pWU7 and the E. coli plasmid pK19 Kmr, which were checked for conjugative transfer from E. coli into the methylotrophic host.

Anti-Bacterial Agents↗

Modification of the Polaron sputter-coater unit for glow-discharge activation of carbon support films.

We describe a modification of the Polaron sputter-coater unit series 11 HD enabling activation of carbon support films for electron microscopy of macromolecules and macromolecular assemblies. The modification is simple and the device can be used in two modes, for sputter-coating of SEM samples and for glow-discharge activation of carbon support films. Examples of protein-free spreading of DNA and negative staining of bacteriophage particles on activated carbon support films are presented.

Bacteriophages↗

Epstein-Barr virus nuclear antigen type 1 binding: electron microscopy.

Epstein-Barr virus (EBV) nuclear antigen type-1 (EBNA-1) was extracted and purified from Raji cells by chromatography on DNA-Sepharose and Blue-dextran Sepharose. Its complexes with plasmid pM765-10 derived from EBV (strain M-ABA) DNA were visualized by electron microscopy. The criteria of specificity were as follows: (1) preferential binding of EBNA-1 to the ori-P region of pM765-10; (2) specific enlargement of EBV DNA/EBNA-1 complexes with anti-EBNA-1 (IR-3) IgG antibody; and (3) resistance of the resulting EBV DNA/EBNA-1/anti-EBNA-1 antibody complexes to treatment with 1.5 M NaCl. The optimal conditions for the formation of EBV DNA/EBNA-1 complexes were 50 to 150 mM NaCl and pH 6.0. A balanced equilibrium of EBNA-1 and pM765-10 was necessary to achieve both a high yield and specificity of EBV DNA/EBNA-1 complexes.

Antigens, Viral↗

New bacteriophage-like particles in Corynebacterium glutamicum.

Three new phage-like particles (CG1, CG2, and CGK1) were isolated from Corynebacterium glutamicum CBII. Particles CG1 and CG2 are DNA phages with long, noncontractile tails, CGK1 is a killer particle according to electron microscopy. A heat-stable low-molecular-weight bacteriocidal substance affecting various coryneform bacteria was observed to be joined to the killer particle CGK1.

Bacteriocins↗

The instability of plasmid pON5300 after its transformation into Escherichia coli K 12.

The plasmid R1drd-19 Km- and its derivative pON5300 were split by the PstI restriction enzyme; they differed in a 2.4 Mg/mol fragment which was present only in the pON5300. Both plasmids were present in 1-2 copies per chromosomal equivalent in E. coli JC5455 cells. The pON5300 is stable in its original mutagenized host but it segregates the enlarged region of the plasmid molecule after transformation into non-mutagenized E. coli JC5455. The possible explanation of this instability is discussed.

Anti-Bacterial Agents↗

Stability of the hybrid plasmid pIM138 and its curing by some eliminating agents.

Hybrid plasmid pIM138 was constructed by insertion of a chromosomal fragment with the threonine operon from Escherichia coli into the pBR322 vector. Molar mass of pIM138 was 2.8 Mg/mol. Heteroduplexes between pBR322 vector and pIM138 hybrid DNA molecules were prepared. The hybrid plasmid shows a high stability against the curing effect of rifampicin and clorobiocin in E. coli SK1590 thr host.

Chromosomes, Bacterial↗

Physical characterization of the plasmid pON5300.

The antibiotic resistance plasmid R1drd19Km- which has spontaneously lost its kanamycin resistance marker, and its derivative pON5300, were analysed using the restriction endonucleases SalI, BamHI, HindIII and EcoRI. The fragment patterns were compared with that of the R1drd19 and the fragments responsible for kanamycin resistance were found to be missing in R1drd19Km- and pON5300. In these plasmids a 7 Mg/mol EcoRI fragment was observed instead of the D (6.3 Mg/mol) fragment of R1drd19. Further a new 6 Mg/mol EcoRI restriction fragment was observed in pON5300. Using double digestions it was shown that the new fragment does not carry restriction sites for HindIII, BamHI and SalI endonucleases. The non-homology of the analysed plasmid was proved electron microscopically by heteroduplex techniques. The possibility of amplification in the regulatory region for the expression of R-determinants in pON5300 is discussed.

DNA Restriction Enzymes↗