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Biomedical subjects

A P Ryskov

Publications and source records attributed to A P Ryskov.

At least 19 recordsLinked to original sources

DNA fingerprints of Bombyx mori L. Testing of genotypic variability of parthenogenetic strains.

A comparative analysis of the total cellular DNA in certain parthenogenetic specimens of the silkworm (Bombyx mori), produced from the females of the parthenogenetic strains by two types of parthenogenesis, has been performed through the application of the DNA fingerprinting method based on M13 phage DNA as a hybridization probe. It has been shown that parental specimens and their genetically identical off-springs produced through ameiotic parthenogenesis have identical patterns of hybridization with the hypervariable DNA fragments. The off-springs produced through the meiotic type of parthenogenesis have individual-specific patterns of hybridization, revealing a high level of polymorphism of individual genotypes. The results obtained testify to the effectiveness and reliability of this promising method for identification of genotypic variability, marking and genomic characterization of parthenogenetic clones in the silkworm.

Animals

[The genomic fingerprinting of the causative agents of sapronoses].

The genome polymorphism of the causative agents of sapronoses (Vibrio cholerae, Legionella and Leptospira) has been studied. The use of the method of genome fingerprinting [correction of dactyloscopy] has been shown to permit the differentiation of closely related strains of such causative agents. The epidemically significant strains of the causative agents of sapronoses, isolated in different geographical regions, have been found to be genotypically related, i.e., they are probably of clonal origin. Avirulent and nontoxigenic strains are genotypically heterogeneous and differ both from one another and from epidemically significant strains. Using V. cholerae as an example, the hypothesis of the appearance of potentially dangerous variants at the epidemic period in the absence of their release at the period between epidemics is considered.

DNA Fingerprinting

[DNA fingerprinting of Rattus norvegicus: a new approach in genetic analysis].

Recent finding in highly effective DNA probes for RFLP testing (of hypervariable minisatellite DNA type) has led to the invention of DNA fingerprinting--the new technique of great value for identification of individuals, establishing biological kinship and studies in population genetics. We anticipate that DNA fingerprinting procedure with M13 phage DNA as a probe which we have developed earlier, makes it possible to apply new approach in genetic analysis--establishing, whether or not a particular locus is associated with the inheritance of genetic disease, by comparing the whole restriction fragment data from affected and unaffected animals. In this work, using the method described we characterized the Kroushynsky-Molodkina rat strain with hereditary disposition for epileptic attacks and performed comparative fingerprint analysis of these defective and normal rat genomes. The data obtained may hold some promises for further seeking the particular defective gene.

Animals

[Analysis of sequences from human brain cDNA gene bank which are functionally active in nervous tissue and tumor cells].

Construction of a human cortex cDNA bank is described as well as the isolation from this bank of pBH71 and pBH3 clones with preferential expression in nervous and in tumor cells. The clones can be included into the third class of cDNA according to Sutcliff's classification. The mRNA corresponding to this cDNA class is considered to play the key role in determination of specificity of nervous tissue. Expression of the pBH71 sequence was revealed in human cortex and in tissues of different genesis (from neuroblastoma to uterus myoma), a 2 kb mRNA which corresponds to one and the same cDNA chain having been found in all tissues under analysis. The nucleotide sequence of cDNA insertion into the pBH71 clone of 447 n.p. was determined, and particular features of cDNA nucleotide composition and possible schemes of its translation were analysed. Weak homology was found between the 3'-end of cDNA insertion of the pBH71 clone and the 3'-end region of human proopiomelanocortine. The cDNA of the pBH3 clone hybridizes with the 0.8 kb mRNA revealed in human cortex and neuroendocrine tumors of different nature. No homology was revealed between the cDNA sequence of the pBH3 clone and any genes deciphered.

Base Sequence

[Study of polymorphism of strains of Vibrio cholerae of various origins using a method of genomic fingerprinting].

Population of atoxigenic Eltor vibrios (vct-) isolated from open water has been shown to be heterogeneous by the genomic fingerprinting technique. Epidemically dangerous strains (vct+) containing the gene coding for cholera toxin have similar hybridization profile independent of time and place of their isolation. The clonal formation of vct+ strains and the absence of vct----vct+ transition of strains under natural conditions are discussed.

DNA, Bacterial

M13 phage DNA as a universal marker for DNA fingerprinting of animals, plants and microorganisms.

Hypervariable polymorphic patterns were detected with M13 phage DNA as a probe in genomic DNA of organisms belonging to different taxonomic groups including animals (vertebrates and invertebrates), plants and microorganisms. Individual-specific restriction pattern analysis (DNA fingerprinting) with this probe proved to be useful for individual identification, analysis of somatic stability and paternity testing in man. The nuclear type of inheritance indicates that the hypervariable DNA regions in question are located in the chromosomes, not in the mitochondrial DNA. The data obtained also demonstrate a potential range of M13 DNA applications as a probe for DNA fingerprinting of animals, plants and microorganisms, particularly for the determination of inbred lines, identification of bacterial strains and establishing stock, variety and strain distinctions.

Animals

[Genomic fingerprinting of microorganisms: its use as a hybridization probe of phage M13 DNA].

Hypervariable nucleotide sequences detected by hybridization with the phage M13 DNA probe were found in the chromosomal DNAs of certain pathogenic microbial species. DNA fingerprinting, based on hybridization of M13-probe with hypervariable chromosomal DNA sequences, opens new approaches to epidemiological analysis, epidemiological prognosis, taxonomy, and other theoretical and applied fields of bacteriology.

Bacteriophages

[Genomic fingerprints of organisms from different taxonomic groups: the use of phage M13 DNA as a hybridization probe].

Hypervariable polymorphic patterns were detected using wild-type M13 DNA as a probe in genomic DNAs of very different organisms ranging from procaryotes and lower eucaryotes to upper plants and animals, including human beings. Due to somatic stability of highly polymorphic patterns and their discrete inheritance, individual-specific restriction pattern analysis ("DNA fingerprinting") with this test probe was found to be useful in applied human genetics, in particular, for identifying paternity and maternity, and mapping of human genomes. The data obtained also demonstrate some possibilities of the DNA fingerprinting technology in genetics and selection of agricultural plants and animals, such as variety analysis, classification and registration of individual inbred lines and strains, as well as identification of bacterial strains.

Animals

Cloning of Alu-containing cDNAs from human fibroblasts and identification of small Alu+ poly(A)+ RNAs in a variety of human normal and tumor cells.

Two clones have been selected from a human fibroblast cDNA bank. By DNA sequencing the clones were shown to contain Alu elements located near the ends of the cDNA inserts. DNA of the clones was used for Northern blot hybridization analysis of a number of poly(A)-containing RNAs from normal human tissues (brain, stomach, uterus, spleen, fibroblasts) and tumors (neurinoma, glioma, neuroblastoma, liposarcoma, adrenal cortex adenocarcinoma). All RNA samples reveal a heterodisperse distribution of Alu transcripts with discrete bands in the region of 7-12 S RNA. The majority of these small poly(A)+ Alu+ RNAs contain Alu sequences only in one (canonical) orientation with functional signals including the split promoter for RNA polymerase III.

Base Sequence

[Expression of genome repetitive sequences in rat bone marrow cells: cloning and characteristics of cDNA-copies of L1 element transcripts].

Recombinant cDNA clones containing the repetitive sequences of major retroposon families--ID, B2 and L1--were isolated from the cDNA bank of rat brain. Their organization was studied by sequencing and hybridization analysis. It was shown for L1 that: 1) its transcripts are mainly found in the fraction of poly(A) +mRNA; 2) there is a population of full-length elements present in the rat genome. We also tried to estimate the copy number of these elements. Some aspects of their organization are discussed.

Animals

DNA sequences homologous to long double-stranded RNA. Transcription of intracisternal A-particle genes and major long repeat of the mouse genome.

Long double-stranded RNA (dsRNA-A) (Kramerov, D.A., Ryskov, A.P. and Georgiev, G.P. (1977) Biochim. Biophys. Acta 475, 461-475) from Ehrlich ascites carcinoma cells was used for the search for mobile dispersed genes in the mouse genome. Two kinds of genomic sequences hybridizing to dsRNA-A were cloned. They were designated A1 and A2. The A1 sequence was identified as the gene for intracisternal A particles, while the A2 sequence was found to be the major long repetitive sequence of the mouse genome. Melted dsRNA-A hybridized equally well to both DNA strands of A1 and A2 sequences while total poly(A)+ RNA bound preferentially to one of them. Thus, a partially symmetrical transcription took place in the case of A1 and A2 elements. The analysis of transcripts of A1 elements in Ehrlich carcinoma cells revealed RNAs with sizes of 9.5 kb, 6.8 kb and 6.0 kb. In plasmocytoma MOPC 21 cells, instead of the 6.0 kb RNA, two other kinds of RNA with sizes 5.3 kb and 7.8 kb were found. These transcripts poorly coincided with the four known variants of intracisternal A-particle (IAP) genes. It seemed that at least some of the described RNAs were transcribed from some minor non-identified variants of IAP genes. The A2 transcripts were practically restricted to nuclei, their sizes being heterogeneous.

Animals

Nucleotide sequence of small polyadenylated B2 RNA.

Small poly(A)-containing RNA molecules which hybridize to the ubiquitous short repetitive sequence B2 and which are transcribed by RNA polymerase III have been identified in the cytoplasm of mouse cells. Here, we describe the structure of this small B2 RNA. A cDNA library was prepared from low-molecular-weight cytoplasmic poly(A)+RNA isolated from Ehrlich carcinoma cells and the clones which hybridized to B2 sequence were selected. The clones were sequenced and shown to contain B2 sequences followed by a poly(A) tract. The sequences of the cloned B2 RNAs different from each other by 3-10%, being similar in this respect to genomic B2 copies. Thus, B2 RNA is transcribed from many different B2 sequences in the genome. The 5'-ends of B2 RNA at least in most molecules coincide with the beginning of B2 genomic sequence. The poly(A) segments located at the 3'-end of small B2 RNA are the same size as in mRNA molecules, suggesting posttranscriptional formation. In some clones, additional sequences were detected between the 3'-end of B2 sequence and the poly(A) stretch. They seem to result from a lesion in the RNA polymerase III terminator in the corresponding B2 sequences. The possible significance of B2 sequences and small B2 RNA is discussed.

Animals