Biomedical subjects
Publications and source records attributed to V P Shchipkov.
The ability of standard plasmids of six Fin-groups to inhibit the functions of genes transferring derepressed F-like plasmids has been studied. It is shown that transposons incorporation into the structure of individual plasmids alters the regulatory system of plasmid tra-genes.
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The data are reviewed on the genetic structure and regulation of tra-genes activity controlling the conjugative transfer of F, F-like plasmids as well as the transfer of some other bacterial plasmids. The effect of the systems inhibiting the conjugational transfer (fin-systems) of F and a number of derepressed F-like plasmids has been characterized on the basis of data obtained by the authors or published data. Possible mechanisms for the systems functioning in the regulation of tra-genes activity are discussed as well as the prospects of their further study.
A study was made of the ability of reference plasmids of the 6 known Fin-groups to inhibit the functions of transfer genes (tra-genes) of the 4 derepressed F-like plasmids (pAP22-2, pAP38, pAP43, pAP53). It was shown that unlike the derepressed Flac plasmid, the conjugation transfer of pAP38 and pAP53 plasmids was inhibited only by, the FinV plasmid, whereas pAP22-2 plasmids by Fin V and Fin V plasmids. The formation of donor-specific pili in case of pAP38 plasmid was inhibited by Fin Q, Fin U and Fin V plasmids, in case of pAP43 plasmid by Fin U Fin V and Fin W plasmids.
Sensitivity of the genetic transfer system of F-like plasmid pAP42 marked with the transposons Tn1 and TN9 to fertility inhibitors of six reference Fin-groups was studied. It was shown that transfer function and donor-specific piliformation of the plasmid under study were inhibited by reference plasmids of FinU and FinV groups, surface exclusion by plasmids of FinU and FinQ groups. The different influence of the FinOP group plasmid on transfer functions of the marked plasmids pAP42::Tn1 and pAP42::Tn9 that is likely to be connected with the effect of incorporated transposons was determined.
F-like pAP19-1 Col-plasmid was labeled with transposons Tn1 and Tn9 and transfer functions of its derepressed mutants were investigated. The plasmid indicated was compatible with reference plasmids of 9 F-like incompatibility groups. Thus it belongs to the new incompatibility group FX. The ability of Tn9 to change the incompatibility of the plasmid investigated was discovered.
It has been demonstrated during investigation of Colplasmid pAP11-2 and its varieties labeled with transpozone (Tn1 and Tn9) that this plasmid is a derepressed one in terms of transfer functions in E. coli strain K-12 cells as well as in some of serologically typed strains of this type. The plasmid under study is incompatible with reference plasmids belonging to two different groups (FI and FIV) and is marked by a number of the properties common to the system of genetic control over Tra-functions.
A study was made of plasmid pAP53 derepressed as regards transfer functions (Tra-functions) detected in E. coli strain cells, serogroup 0128, after its labeling with transpozones Tn1 and Tn9. The compatibility tests demonstrated that the plasmid belongs to the incompatibility group FIII and is partially incompatible with the group FII reference-plasmid. Plasmid pAP53 is unable to inhibit Tra-functions of plasmid F'lac and is not inhibited by the fin type genetic regulation on the OP group plasmids under study. At the same time Tra-functions of plasmid pAP53 are inhibited in the presence of pAP41 plasmid, which indicates that this plasmid has a special type of genetic regulation.
A group of conjugation plasmids identified in the cells of wild strains of E. coli isolated from the people and animals were examined. The majority of plasmids showed the ability to inhibit Flac plasmid genetic transfer functions. However, some of them differed in the levels of inhibitory activity. Similar experiments with derepressed F-like pAP22-4 plasmid demonstrated that it is marked by a less ability (as compared with Flac plasmid) to be inhibited by different plasmids. This circumstance is possibly related to the specificity of the genetic regulation of plasmid transfer functions.
A study was made of the genetic traits of F-like plasmid pAP10-2 that monitors the synthesis of thermostable enterotoxin after plasmid labelling with Tn9 transpozone. It has been shown that the test Ent-plasmid is a conjugative one, suppresses fertility functions of F plasmid and belongs to the F1 incompatibility group.
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The study on 63 strains of conditionally pathogenic E. coli isolated from patients with chronic pyelonephritis showed that the bacteria of the 47 strains (74.6%) were resistant to one or several antibiotics. 7 strains of such bacteria carried R-plasmids capable of transferring into the cells of E. coli K-12 during conjugation. Most of the identified 9 R-plasmids belonged to the class of the F-like plasmids and were characterized by similar lots of antibiotic resistance markers. DNA of the detected R-plasmids were treated with endonuclease (restrictase) EcoRI. After that the fragments were analysed by means of electrophoresis in agarose gel. On the basis of the restrictogrames it was concluded that the plasmids studied were not identical. A possibility of detecting complexes consisting of different plasmids carrying similar lots of antibiotic resistance markers in the cells of the same strain was shown.
A number of conjugative plasmids of opportunistic E. coli with molecular weights of 23.6--81.8 megadaltons have been studied. On the basis of the data thus obtained conjugative F-like R-plastmids have been subdivided into 2 groups characterized by the unsuppressed or suppressed manifestation of their functions. Individual conjugative plasmids have been found to differ in their capacity for mobilizing non-conjugative SuSm plasmid for transfer, which indicates the possibility of using this test for characterization and classification of plasmids, identified in natural bacterial population.
The plasmid DNAs isolated from E. coli AP1 were studied by electron microscopy. Two plasmid DNA forms (FB1-1 and FB1--2) with the mol wt of 35.9 +/- 0.5 x x 10(6) and 51.5 +/- 0.6 x 10(6) daltons, respectively, were revealed.
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Compatibility of depressed F-like plasmid FB1drd integrated into E. coli K12 cell chromosome with standard plasmids of FI-FVI compatibility groups was studied. The results obtained show that such plasmids can stably coexist in the same cell with plasmid FB1drd. On these grounds it is supposed that the latter belongs to the new compatibility group (FVII).
F-like plasmids (FBl, FBl drd, F-lac+) were transferred to the recipient strains E. coli, K-12 with different recombination ability. Then the sensitivity of these strains to ultra-violet irradiation and also the stability of the mentioned plasmids under conditions of bacteria treatment with elimination agents was determined. The presence of any F-like plasmids under study failed to influence the ultra-violet sensitivity of the bacterial cells. On the other hand, the research results of spontaneous and inducec elimination of these plasmids indicated considerable differences in relation to the elimination agents. It is supposed that the stability of individual F-like plasmids in the cells largely depended on the genetic differences in rec A gene in the bacterial cells containing them.
The authors studied 130 strains of serologically typed E. coli for the capacity to provide reproduction of donor-specific phages MS2, Qbeta, and f1; positive increase of the donor-specific phages reaction was found in 12 cases. Some of the detected L-like plasmides bore determinants of colicinogenicity or drug resistance. Such plasmides were transmissive and capable of inhibiting donor properties controlled by factor F'-lac+, i.e. they served as plasmides of type fi+.