[Rhizosphere strain of Pseudomonas chlororaphis capable of degrading naphthalene in the presence of cobalt/nickel].

Combination of genetic systems of degradation of polyaromatic hydrocarbons, resistance to heavy metals, and promotion of plant growth/protection is one of the approaches to the creation of polyfunctional strains for phytoremediation of soils after combined contamination with organic pollutants and heavy metals. A plant-growth-promoting rhizosphere strain Pseudomonas chlororaphis PCL1391 (pBS216*, pBS501) has been obtained, in which the nah operon of plasmid pBS216 provides naphthalene biodegradation and the cnr-like operon of plasmid pBS501 provides resistance to cobalt and nickel due to the withdrawal of heavy metal cations from the cells. In the presence of 100 microM of nickel, the viability, growth rate, and naphthalene biodegradation efficiency of the resistant strain PCL1391 (pBS216*, pBS501) were much higher as compared with the sensitive PCL1391 (pBS216). During the growth of the resistant strain, in contrast to the sensitive strain, nickel (100 microM) had no inhibiting effect on the activity of the key enzymes of naphthalene biodegradation.

[PCR-based diagnostics of community-acquired pneumonia of mycoplasmal and Chlamydia etiologies].

Two hundred and ninety-two patients with pneumonia evidenced by clinical, X-ray and laboratory findings were examined for the purpose of studying the etiological significance of Mycoplasma pneumoniae and Chlamydophila pneumoniae--atypical pathogens; the patients, members of closed bodies, were admitted to the hospital for as long as one year. PCR was used to examine the sputum of patients who were at exacerbation. It was established that the mentioned pathogens prevailed in autumn and winter. It can be concluded, on the basis of a PCR-based monitoring of the atypical pathogens, that the significance of the atypical pathogens within the etiological structure of pneumonia acquired in isolated collective by young persons has been on the rise.

[Detection of Mycoplasma pneumoniae and Chlamydophila pneumoniae in patients with community acquired pneumonia].

A total of 292 patients with pneumonia confirmed by clinical, roentgenological and laboratory methods, admitted to the hospital from closed communities during the one year period, were examined. The sputa of patients with pneumonia in the acute stage were studied in the polymerase chain reaction (PCR). The predominence of M. pneumoniae and C. pneumoniae causualties in autumn and winter period was established. The monitoring of atypical infective agents with the use of the PCR techniques gives evidence for conclusion on their ever growing role in the etiological structure of community acquired pneumonia in groups of closely interacting young people.

[Anaerobic degradation of biphenyl by the facultative anaerobic strain Citrobacter freundi BS2211]. / Anaérobnaia degradatsiia bifenila fakul'tativno anaérobnym shtammom Citrobacter freundii BS2211.

Using a synthetic medium supplemented with biphenyl (a polycyclic aromatic hydrocarbon), a new bacterial strain of Citrobacter freundii was isolated from enrichment cultures containing soil and industrial wastewater samples of the Serpukhov Condenser Factory. This strain was found to be capable of degrading biphenyl under anaerobic conditions in the course of nitrate reduction. When the initial concentration of biphenyl in culture medium equaled 150 mg/l, the culture with a titer of 10(9) cells/ml degraded up to 26-28% of biphenyl in 3 days (28 degrees C). At 250 mg/l, the culture with a titer of 10(7) cells/ml degraded 15% of biphenyl in 21 days. Approximately 10% of the substrate consumed was utilized completely, whereas the remainder underwent transformation.

[Isolation and characteristics of an ordered library of transcribed sequences of human chromosome 19 from hybrid human-hamster cells]. / Poluchenie i kharakteristiki uporiadochennoi biblioteki transkribiruemykh posledovatel'nostei khromosomy 19 cheloveka iz gibridnykh kletok chelovek-khomiak.

A new technique based on Alu-PCR amplification of hn-RNA is described for the extraction of human-specific transcribed sequences from a hybrid cell line. Arrayed library of hn-cDNA was constructed and characterized by sequencing about 80 individual clones. A high enrichment by human-specific sequences (about 95%) was demonstrated.

[Construction of a mapping panel of human-rodent hybrid cells]. / Sozdanie kartiruiushchei paneli gibridnykh kletok chelovek-gryzun.

A clone panel of 27 human-Chinese hamster and 4 human-mouse somatic cell hybrids which contained as minimum five discriminating clones for any chromosome pairs was set up. Segregation analysis of 45 human chromosome-specific isoenzymes and PCR markers in hybrid clones allowed to demonstrate a possibility to apply the obtained panel for chromosome mapping of human genes.

[Use of isoenzyme and PCR markers for analyzing human chromosomes in human-rodent hybrid cells]. / Ispol'zovanie izofermentnykh i PCR-markerov dlia analiza khromosom cheloveka v gibridnykh kletkakh chelovek-gryzun.

The possibility of using isozyme and PCR markers for estimation of preservation of human chromosomes in human.rodent somatic cell hybrids is considered. Methods of electrophoretic separation of 33 isozymes and 11 PCR markers for 22 human autosomes and X chromosome are described. Using these isozymes and PCR primers as markers of known regions and arms of human chromosomes, one can avoid errors in typization of chromosomes with complicated rearrangements which simulate similar patterns of G-banding. The method proposed in the paper not only facilitates considerably the analysis of hybrid clones, but is also an easy and reliable tool in selection of somatic cell hybrid clones for further investigation.

[An analysis of the gene coding for the basic neutralizing antigen VP7 of human rotavirus isolate 1407]. / Analiz gena, kodiruiushchego osnovnoi neitralizuiushchii antigen VP7 izoliata 1407 rotavirusa cheloveka.

A procedure based on polymerase chain reaction was developed for the study of rotaviruses. A full-length cDNA copy of the gene coding for major neutralizing glycoprotein VP7 of a human rotavirus isolate 1407 with the "long" electrophoretype (classified as the first electrophoretype) was cloned and sequenced. The primary structure of glycoprotein VP7 of isolate 1407 in A and C antigenic regions was found to be similar to that of serotype 1 virus.

[Use of the polymerase chain reaction for analysis of rotaviruses. Nucleotide sequence of a gene, coding for the basic neutralizing antigen VR7 of a human rotavirus with a new G-serotype]. / Ispol'zovanie polimeraznoi tsepnoi reaktsii dlia analiza rotavirusov. Nukleotidnaia posledovatel'nost' gena, kodiruiushchego osnovnoi neitralizuiushchii antigen VP7 rotavirusa cheloveka novogo G-serotipa.

A procedure based on polymerase chain reaction use for the detection of rotavirus has been developed. Full length cDNA copy of the VP7 gene coding for the major neutralization glycoprotein of the human rotavirus RK9 with an unusual "wide" electrophoretype is cloned and sequenced. Glycoprotein VP7 of RK9 has a unique amino acid composition in A and C antigenic regions. It shows that strain RK9 represents a new (12) rotavirus serotype.

[Rare initiation codons are regulators of expression of the rpoC gene]. / Redkie initsiiruiushchie kodony-reguliatory ékspressii gena rpoC.

Translation of the rpoC genes in Escherichia coli and Salmonella typhimurium is known to start from the GUG codon. Now, using toeprint analysis we have shown UUG to be the initiation codon of the Pseudomonas putida rpoC gene. IF3 does not seem to proofread initiation at the UUG codon. The rpoC genes of P. putida, E. coli, and S. typhimurium, which use rare start codons, have strong SD-domains AGGAGG (P. p.) and GGGAG (E. c., S. t.), optimal seven-nucleotide spacing between SD and start codons, and good second codon AAA. We suggest that rpoC presents an infrequent case of the regulation of translation initiation by selecting the start codon.

[Isolation of the porcine alpha-interferon gene using simultaneous directed multipoint mutagenesis]. / Poluchenie gena alpha-interferona svin'i metodom odnovremennogo napravlennogo mnogotochechnogo mutageneza.

A method of obtaining the pig alpha-interferon gene by means of simultaneous multidirected mutagenesis of the human alpha 2-interferon gene is presented. Nucleotide homology between these genes is 80.4%. Fourteen synthetic oligonucleotides forming a pig alpha-interferon gene's strand were ligated on a single-stranded template, carrying cDNA of the human alpha 2-interferon gene. The obtained DNA fragment was cloned in the single-stranded or double-stranded form. It was found that the method does not affect the cloning efficiency. The primary structure of the gene was confirmed by sequencing.

Effective method for obtaining long nucleotide chains on partially complementary templates. Processed bovine gamma-interferon gene obtained from human gamma-interferon gene.

The method of obtaining the bovine gamma-interferon gene by means of simultaneous multidirected mutagenesis of the human gamma-interferon gene is presented. The first strand of the bovine gamma-interferon gene was obtained by ligation of synthetic oligonucleotides, using the cDNA of human gamma-interferon, cloned in the single-stranded phage M13mp19 as a template. The second strand was synthesized using a large fragment of E. coli DNA-polymerase I. The double-stranded gene was then treated by restriction nucleases and cloned in a pUC-18 derived vector. The primary structure was confirmed by sequencing.

[Nucleotide sequence of the rplL gene coding for ribosomal protein L7/L12 of Pseudomonas putida]. / Nukleotidnaia posledovatel'nost' gena rplL, kodiruiushchego ribosomnyi belok L7/L12 Pseudomonas putida.

The Pseudomonas putida rpl L gene coding for ribosomal protein L7/L12 was cloned and sequenced. Although Asp55 residue in L7/L12 was previously shown to be conservative in ten different organisms, the Pseudomonas putida L7/L12 proved to contain Asn55, thus showing that Asp55 is not invariant.

[Genes coding for RNA polymerase in bacteria. III. The use of modified Sanger's method for sequencing the C-terminal region of rpoB gene, N-terminal region of rpoC gene and intercistron region of RNA polymerase in Pseudomonas putida]. / Geny, kodiruiushchie RNK-polimerazu bakterii. III. Ispol'zovanie modifitsirovannogo metoda Sengera dlia opredeleniia pervichnoi struktury C-kontsevoi oblasti gena rpoB, N-kontsevoi oblasti gena rpoC i mezhtsistronnogo uchastka RNK-polimerazy Pseudomonas putida.

The Sanger method was modified and the primary structure of the SalI-C fragment of the Pseudomonas putida rpoBC operon was elucidated.

[Genes coding for RNA-polymerase in bacteria. II. Conservative sites in the central region of the beta-subunit of Pseudomonas putida RNA-polymerase]. / Geny, kodiruiushchie RNK-polimerazu bakterii. II. Konservativnye uchastki v tsentral'noi chasti beta-sub''edinitsy RNK-polimerazy Pseudomonas putida.

SalI--L fragment of the P. putida rpoBC operon has been sequenced and conservative regions of the central part of the RNA-polymerase beta-subunit have been determined. Amino and acid residues interacting with Zn2+ are postulated.

[Amino acid substitutions in the beta-subunit of RNA-polymerase from E. coli compensating for mutation-induced damage of the rho termination factors]. / Aminokislotnye zameny v beta-sub''edinitse RNK-polimerazy E. coli, kompensiruiushchie mutatsionnye povrezhdeniia faktora terminatsii ro.

Ts-phenotype of the E. coli rho-factor mutant rho 15 is suppressed by two rifampicin-resistance mutations, rhoB1019 resulting in a single amino acid substitution Val146----Phe and rhoB268 resulting in a single substitution Gln513----Leu in beta-subunit of the E. coli RNA polymerase. Rifampicin-resistance mutations rhoB255 (Asp516----Val), rhoB1016 (Asp516----Asn), rhoB1001 (His526----Tyr), rhoB1004 (Ser531----Phe), rhoB1005 (Pro564----Leu), and streptolydigin-resistance' mutation rhoB1018 (double substitution Gly544----Asp and Phe545----Ser) do not suppress the rho15 mutation.

[Genes encoding bacterial RNA-polymerase. I. Cloning of rpoBC-operon of Pseudomonas putida and its physical map]. / Geny, kodiruiushchie RNK-polimerazu bakterii. I. Klonirovanie rpoBC-operona Pseudomonas putida i ego fizicheskaia karta.

The P. putida rpoBC operon, coding for beta and beta' subunits of RNA polymerase, was cloned and its physical map constructed.