[DNA fingerprinting of parthenogenetic families of Lacerta-species lizards: detection of genetically unstable loci].
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Animals↗
Biomedical subjects
V V Grechko
Publications and source records attributed to V V Grechko.
[Reticulate evolution of parthenogenetic species of the Lacertidae rock lizards: inheritance of CLsat tandem repeats and anonymous RAPD markers].
The genetic relatedness of several bisexual and of four unisexual "Lacerta saxicola complex" lizards was studied, using monomer sequences of the complex-specific CLsat tandem repeats and anonymous RAPD markers. Genomes of parthenospecies were shown to include different satellite monomers. The structure of each such monomer is specific for a certain pair of bisexual species. This fact might be interpreted in favor of co-dominant inheritance of these markers in bisexual species hybridogenesis. This idea is supported by the results obtained with RAPD markers; i.e., unisexual species genomes include only the loci characteristic of certain bisexual species. At the same time, in neither case parthenospecies possess specific, autoapomorphic loci that were not present in this or that bisexual species.
Animals↗
[Ribonucleoproteins containing mRNA in the cytoplasm of mouse plasmacytoma cells].
The rapidly labelled postribosomal ribonucleoprotein (RNP) found in the cytoplasm of mouse plasmacytoma cells were investigated. It has been shown that 45S and 80S particles contain relatively high molecular weight (approximately 12-17S) pulse-labelled RNA similar to the polyribosomal mRNA. No other postribosomal RNP was found which would contain an RNA with similar sedimentation characteristics. In CsC1 density gradients, the postribosomal RNP gives two peaks. One of them, the rapidly labelled component (rho 1.52 g/cm3) is found only in 45S RNP. The other rapidly labelled component (rho 1.36-1.41 g/cm3) is revealed in all investigated regions of sucrose gradients. The latter contains relatively low molecular weight RNA (approximately7-9S). These RNP are supposed to be informosome-like particles. The components with a buoyant density of 1.52 g/cm3 may represent an mRNP-45S subparticles complex. The rapidly labelled mRNA of 80S particles is released after EDTA treatment in the form of mRNP with a buoyant density of 1.45-1.47 g/cm3.
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[Taxonometric analysis of DNA repeating elements].
The method of analysis of low molecular weight fragments of high copied repeats of DNA hydrolysed by restriction nuclease is presented. The P labeled fragments (by means of DNA polymerase I) are electrophoresed in nondenaturing PAAG and radio-autographed. The specific band patterns are observed in region between approximately 20 and 300 bp. When studying some lizard and fish species DNA's it was shown that the patterns observed have species and genus specificity but not individual. The approach supposed can be applied to investigation of interspecies relationships, some evolutionary problems and to the studying of questions concerning the role of DNA repeats in evolution. The method is simple and comparatively cheap and may be called as "taxonomic DNA fingerprint" or "DNA taxonoprint" method.
Animals↗
[Molecular markers of nuclear DNA in the study of evolution and speciation process in an example of "Lacerta agilis complex" (Sauria: Lacertidae)].
In order to elucidate the molecular-genetic relations of some Lacerta s. str. lizard populations, subspecies and species in comparison with some other genera we used methods revealing four types of nuclear DNA markers. Among these were taxonprint, RAPD, Inter-MIR-PCR markers and also satellite DNA monomer sequences. The aim was to compare the phylogeny and systematics of this reptilian group based on morphological and molecular criteria. This problem has a general importance for understanding a speciation process. Our results show a good correlation between both approaches when genera and species levels were studied. Systematic status of five subspecies of L. agilis were supported but not in all cases, some subspecies have no meaningful genetic differences by three types of molecular markers, but all of them were differed by RAPD markers. The data confirm the subdivision of L. agilis populations into west and east clades proposed by other authors earlier on the basis of mitochondrial DNA and morphology. The population structure of one of the subspecies--L. agilis exigua, was studied on a number of populations distributed from Ural region up to Kabardino-Balkaria by IMP method. There were no significant differences among these 14 populations investigated. The data testify the rapid distribution of the species after the end of Pleistocene glaciation.
Animals↗
[A new family of interspersed repeats from squamate reptiles].
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Animals↗
[Comparison of the conformation of RNA from phage MS2 and 16S rRNA. Accessibility to nucleases S1 and SV specific to secondary structure and thermal stability].
The hydrolysis of E. coli 16S rRNA by nucleases specific to the secondary structure elements (S1 and SV), the melting of the RNA after partial hydrolysis by nuclease S1 and the electrophoretic mobility of hydrolysis products after denaturation-renaturation of RNA were studied. It was shown that the sensitivity of 16S rRNA to nuclease S1 depends on Zn or Mg ions concentration. The melting curves after partial hydrolysis by nuclease S1 were characterized by a decrease of the hyperchromic effect (by approximately 15%) and by a increase of Tm (by 3 degrees). After RNA denaturation followed by slow or fast renaturation the electrophoretic patterns of the hydrolysis products were not changed, as in the case of phage MS2 RNA. It was supposed, that the rRNA molecule has a stable "nucleus" (or "nuclei"), which is organized as an intramolecular association of parallelly oriented double-stranded fragments of this RNA. Previously, such a mode of the spatial organization was proposed by us for phage MS2 RNA.
Coliphages↗
[Comparison of the conformation of RNA from phage MS2 and 16S rRNA. Interaction with dyes specific for the secondary structure of native RNA and RNA subjected to hydrolysis by nuclease S1].
The interaction of ethidium bromide (EtBr) with double-stranded (ds), and acridine orange (AO) with single-stranded (ss) fragments of 16S rRNA Escherichia coli in a wide range of ionic strength, at various pH, Zn2+ ion concentrations and partial hydrolysis by nuclease S1 was investigated. It was shown that about 90% of the RNA molecule is accessible to both dyes, when the ionic strength is near of 0.01 (pH 7). Approximately half of the RNA becomes inaccessible to dyes, when the ionic strength was increased up to 0.08-0.24 (pH 4.7-7), independent on the presence of Zn2+ ions (10(-3) M). About a half of the ds-, and a quarter of the ss-segments of the RNA, deduced from the secondary structure model were protected from the interaction with EtBr and AO. The hydrolysis of about a half of ss-segments upon addition of the Zn2+ (10(-3) M) ions did not affect the RNA tertiary structure. The experimental data obtained confirm the idea of the existence of some "nucleus" (or "nuclei") within the 16S rRNA molecule. The "nucleus" seems to be inaccessible to the dyes and is very stable to heat denaturation. It was supposed that this structure is organized by means of interaction of some of the parallelly oriented ds-segments, as it was suggested earlier for the phage MS2 RNA structure.
Acridine Orange↗
[Cytoplasmic ribonucleotides of the cells of the rabbit spleen].
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Animals↗
[Formation of C-14-alanyl-RNA].
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Alanine↗
[Study of the secondary and tertiary structure of phage MS2 RNA using nucleases and fluorescent dyes specific for secondary structure].
The binding of ethidium bromide (EtBr) and acridine orange (AO) to RNA in native state or after hydrolysis by S1 and SV nucleases that specifically split single-stranded and double-stranded segments was studied. Nuclease S1 hydrolysis of RNA does not increase the number of EtBr strong binding sites, Tm and hyperchromic effect being also unchanged. Hydrolysis by double-stranded segments accessible to EtBr is followed by the diminishing of Tm and hyperchromism. A supposition is put forward that the main role in stabilization of the RNA tertiary structure is played by double-stranded segments arranged so that some of them are hidden and do not interact with dyes. One of the possible models may be parallel oriented intramolecular "hair-pins" forming compact "rod-like" structures.
Coliphages↗
[Isolation and characterization of the matrix properties of electrophoretically homogeneous immunoglobulin L-chain mRNA from mouse myeloma MORC 21].
mRNA with m. w. 4.10(5) dalton was isolated by elution of one of the fractions of poly(A)-RNA from polyacrylamide gel. During reelectrophoresis this mRNA was electrophoretically homogeneous and had the same size. The isolated fraction represented the template activity in a wheat germ cell-free system of protein synthesis and stimulated the synthesis of polypeptides, electrophoretically correspondent to L-chain of authentic immunoglobulin. Comparative data of properties of electrophoretically homogenous mRNA and analogous fraction of mRNA, isolated from sucrose concentration gradient after fractionation of total poly(A)-containing RNA are given.
Animals↗
[Synthesis of double-stranded DNA on light immunoglobulin chain matrix RNA].
cDNA was synthesized on Ig kappa-L-chain mRNA isolated from mouse plasmocytoma MOPC 21 using avial myeloblastosis RNA-dependent DNA polymerase. The cDNA was used as a matrix for second DNA chain synthesis using 5'-exonuclease free DNA-polymerase I (Klenoff fragment). Hairpin DNA having double length of initial cDNA was obtained without added exogenous primer. SI nuclease treatment of the hairpin DNA results in reducing the DNA length twice, as shown by electrophoresis in denaturing conditions. The fact, that S1 nuclease resistance of the hairpin DNA is 75% shows high complementarity between first and second DNA chain. The product length obtained after S1 nuclease treatment was shown to be heterogenous. The maximal length of the product, reaching at least 900 base pairs, is near to be initial mRNA length (without the poly(A)-fragment).
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[Study of spatial organization of the RNA of phage MSZ using nucleases specific for the secondary structure].
The limited hydrolisis of bacteriophage MS2 RNA by nuclease S1 and ds-specific snake venom RNase was studied in a wide range of ionic strength, at different pH, after heating and (slow and fast) cooling and at various enzyme-substrate relations. It was shown that the RNA has exposed hydrolisis sites for both nucleases. The localizations of these sites are very specific and are not altered in all conditions studied. The hydrolisis rate was changed in some conditions, at that the fragments patterns in denaturing electrophoresis did not move. It was supposed that the RNA has strongly predetermined and predominant conformation which could not be altered by strong influences.
Coliphages↗
[Study of the structural organization of RNA from phage MS2 using fluorescent dyes].
The spatial organization of phage MS2 RNA by binding to ethidium bromide (EtBr) and acridine orange (AO) to RNA was studied. The analyses of dye interaction by spectrophotometric and fluorometric methods have demonstrated that only about a half of 65-70% nucleotides of double-stranded segments can interact with AO and EtBr. On the other hand all the single-stranded segments appear to be accessible to AO binding. These interactions did not practically change when ionic strength (0.01-0.3), Mg2+ and Zn2+ concentrations (10(-3) M) or pH (4.7-7.4) varied. The data permit to suppose that phage MS2 RNA has a very stable tertiary structure which makes part of double-stranded segments unaccessible to inter calating dyes. Taking these and other facts into consideration we suppose that double-stranded segments play an important role in stabilization of the RNA tertiary structure. One of the most possible structure is a compact "rod-like" intramolecular aggregate of double-stranded hairpin-like segments of RNA with parallel orientation.
Coliphages↗
[Translation of MOPC 21 plasmacytoma cell mRNA in a cell-free wheat germ system].
A wheat germ cell-free system was used for translation of kappa-chain IgG1 messenger RNA isolated from mouse plasmacytoma MOPC 21 cells polysomes. The system was optimised for a number of ingredients of the incubation mixture. Incorporation of labelled amino acids in polypeptides was shown as a function of K+, Mg2+, exogenous mRNA concentration and time, mRNA was purificated by two successive oligo (dT)-cellulose chromatographies and two successive sucrose gradient centrifugations or polyacrylamide gel electrophoresis. Different fractions of poly(A)-RNA stimulated protein synthesis 20--30-fold in a wheat germ cell-free system. Analysis of the translation product of 14 and 16S mRNA by SDS-polyacrylamide gel electrophoresis revealed polypeptide comigrating with the authentic L-chain IgG1 and specifically precipitating by an antiserum to mouse IgG and L-chain IgGl (MOPC 21). The 18 and 28S mRNA fractions directed the synthesis of polypeptides with molecular weight up to 50 000 dalton. Immunospecific translation product of the 18 and 28S mRNA contain both L- and H-chains.
Animals↗
[Taxonoprint DNA analysis of certain lizard species from the Lacertidae family].
It was earlier shown that electrophoretic patterns of the highly repetitive segments of DNA's of some animal species studied seem to have species specificity ("DNA taxonoprint method") [2]. In this paper the results of investigation of more than 20 lizard species of the Lacerta genus in comparison with representatives of some other genera and family are presented. It was shown that in taxonoprints there are one band, at least, but usually more bands common for all Lacerta species. Specimens of genus Podarcis have more similarity with genus Lacerta than genus Eremias. The representatives of Teiidae familie have no resemblance with family Lacertidae at all. Among Lacerta species there are 10, which form the separate group being very similar by most of the taxonoprints. This fact goes in line with hypothesis that 5 parthenogenetic species of this group evolve due to interspecies hybridization of another 5 bisexual species of this group. The other species studied have specific peculiarities in various taxonoprints permitting to draw some conclusions on their relations.
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[Study of the relationship of parthenogenetic forms of lizards of the species Lacerta and proposed parental bisexual species in comparison of DNA segments, considered by the polymerase chain reaction with a single primer].
Due to some suggestion [8] the parthenogenesis in vertebrates is the result of interspecies hybridization, and some species seem to be the main candidates participating in this process. DNA parts, which may be revealed using polymerase chain reaction in AP-RAPD modification, were compared in parthenogenetic lizards forms and in supposed parental species for them. As it was shown by other authors and confirmed by us, arbitrary primed randomly amplified DNA products are species specific and may be used as a molecular taxonomic criterion. It was shown that each parthenogenetic species studied has the patterns containing only the bands of one or another supposed parents and no more. This is in favour of the hypothesis mentioned above. Investigation of 103 10-nucleotide primers in PCR was done.
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