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Biomedical subjects

A S Grafodatskiĭ

Publications and source records attributed to A S Grafodatskiĭ.

At least 19 recordsLinked to original sources

[Comparative chromosome painting].

The review of the data on comparative chromosomal painting in mammals is presented. The development of new molecular-cytogenetic methods has resulted in the accumulation of the detailed information on homology of chromosomal segments of more than 50 species from 11 orders. In this review, modern methods of obtaining painting probes are considered in detail, and the basic tendencies of karyotype evolution in different taxa are discussed. Putative karyotypes of the ancestors of primates, carnivores, and placental mammals are considered.

Animals↗

[A new approach to the analysis of complex chromosomal rearrangements in cell hybrids].

The chromosomal complements of somatic cell pig-mink hybrids was determined by a new approach. This approach includes microdissection of metaphase chromosomes, generation of chromosome and region-specific DNA libraries, and fluorescence in situ hybridization of these libraries with pig lymphocyte chromosomes. The studied hybrid cells were shown to contain two small acrocentric chromosomes and a microchromosome of porcine origin. Identification of these chromosomes by differential GTG-staining was impossible. Chromosome isolation by a micromanipulation technique followed by DNA amplification in TOPO-DOP polymerase chain reaction provided chromosome-specific DNA libraries of the rearranged chromosomes. Based on these libraries, the labeled DNA probes were prepared and hybridized to pig chromosomes. This allowed us to determine the origin of the material contributing to the hybrid cell chromosomes. One of these chromosomes contained five pig chromosomal regions: 15cen-q2; 6q21-q23; 13q21; 13q22; 7q25-qter, while the other contained the following pig chromosomal regions: 4p12-p13; 16q12-q14; 12pter-p15. The microchromosome contained the Xp11-Xq11 region. The minimal size of the revealed chromosomal regions was about 3 to 4 x 10(6) bp. Segregation analysis of the thymidine kinase gene 1 (TK1), which was earlier localized to the pig 12p region, and the hybrid cell pig chromosomes in the hybrid subclones suggested that TK1 gene can be assigned to 12p15-pter. The results obtained demonstrate the efficiency of the applied approach in its detailed and reliable description of complex chromosomal rearrangements in hybrid clones, when differential chromosome staining failed to identify these chromosomes.

Animals↗

[Cosmid libraries containing DNA from human chromosome 13].

We characterized two cosmid libraries constructed from flow-sorted chromosome 13 at the Imperial Cancer Research Fund (ICRF), UK (13,000 clones) and Los Alamos National Laboratory (LANL), USA (17,000 clones). After storage for two years, clones showed high viability (95%) and structural stability. EcoR I and Hind III restriction patterns were studied in more than 500 ICRF and 200 LANL cosmids. The average size of inserts was shown to be 35-37 kb in both the libraries. Most cosmids (83% and 93% of ICRF and LANL libraries, respectively) exceed the lower size limit of DNA fragments that can be packaged and represent a good source for physical mapping of chromosome 13. Total length of inserts is four and five genome equivalents in the ICRF and LANL libraries, respectively. ICRF cosmids showed hybridization to 22 of 24 unique probes tested, which corresponds to a 90% probability of having any DNA fragment represented in the library. More than 1 Mb of chromosome 13 is overlapped by 90 cosmids of 22 groups revealed. A chromosomal region of more than 150 kb, containing the ATP1AL1 gene for alpha-1 peptide of Na+, K(+)-ATPase, is covered by 12 cosmids forming a contig. The results of restriction and hybridization analyses are stored in a CLONE database. These data and all the cosmids described are publicly available.

Chromosome Mapping↗

[EcoRI- and BamHI-families of repeated sequences in mustelids].

The restriction enzymes EcoRI and BamHI digest the genomic DNAs from six mustelids species Mustela lutreola, M. vision, M. erminea, M. sibirica, Vormela peregusna, producing repeated fragments varying in length. Some fragments were hybridized to chromosomes and restriction digests of DNAs from some mustelids and other mammals. The 0.7 kb EcoRI repeats from DNA of M. erminea are dispersed over chromosomes of carnivors. The 1.35, 1.9 and 2.7 kb BamHI repeats from DNA of polecat M. putorius furo are specific for mustelids. These repeats demonstrate interspecific variation in length and the number of copies. All BamHI repeats have no strict tandem organization. The 1.9 kb BamHI repeats are concentrated in the heterochromatic pericentromeric regions and additional chromosome arms. The 1.35 kb BamHI repeats are only located in the centromeric regions of chromosomes of five species and are absent in Vormela peregusna.

Animals↗

[Genome mapping in silver fox. Syntenic genes in Carnivora].

Hamster X fox somatic cell hybrids segregating individual fox chromosomes in different combinations were used to assign seven structural loci to fox chromosomes. The gene for ME1 was mapped on the VFU1 chromosome, the genes for ADK and PP being located on the VFU4 chromosome. The gene for GSR was assigned to the VFU7 chromosome and the genes for MPI and COT1 were assigned to the VFU15 chromosome. Localization of these genes enhances the established fox genetic map and extends the known syntenic homologies between the fox and other mammalian. The comparison of data on gene mapping has provided basis for suggestion that there are significant differences in rates of karyotypic evolution in many mammalian taxa.

Animals↗

[Phylogenetic relations of closely related species of Mustelidae. Interspecies variability of localization of restriction sites of BamHI-repeats].

The patterns of blot-hybridization of cloned BamHI repeats to genome DNAs were applied for estimation of phylogenetic relationships of closely related species (Mustela (L.) lutreola, M. (P.) putorius, M. (K.) sibirica, M. (M.) erminea, M. (L.) vision, Vormela peregusna). On the basis of information about interspecific distribution of the blot-hybridization bands (+, -) of BamHI repeats, phylogenetic tree was constructed, after selection of compatible characters, which revealed essential rate ununiformity during mustelids' evolution.

Animals↗

[Genome mapping of the silver fox. I. Determination of the chromosomal location of 8 fox genes and the search for homologous regions on fox and human chromosomes].

Twenty-three silver fox-Chinese hamster somatic cell hybrids were analysed for the expression of fox enzyme loci and the segregation of fox chromosomes. This analysis made it possible to assign the gene PGD to chromosome 2, MDH2 to chromosome 3. NP to chromosome 10. APRT, ENO1, PGM1 to chromosome 12, MDH1 and IDH1 to chromosome 16. Possible use of the above-mentioned clone panel for fox gene mapping is analysed. An attempt to reveal homologous regions on fox and human chromosomes was made by comparative analysis of prometaphase fox and human chromosomes containing the homologous genes. The means and perspectives of verification of the hypothesis proposed are discussed.

Animals↗

[Karyotypes of 5 species of Soricidae-Crocidura in the fauna of the USSR].

The chromosomes of five Crocidura species have been studied by means of routine, G-, C- and Ag NOR-banding techniques. The karyotypes of the species with 2n = 40, i.e. C. gueldenstaedtii, C. suaveolens and C. sibirica, are identical. Short arms of the Y-chromosome of C. sibirica are larger. The karyotypes of C. leucodon (2n = 28) and C. zarudnyi (2n = 22) could be obtained as a result of tandem and centric fusion and pericentric inversions. Small amounts of heterochromatin were detected in autosomes of the species investigated. The Y-chromosomes are completely heterochromatic. Four NOR-bearing pairs are detected in the species investigated.

Animals↗

[Chromosomal localization and evolutionary age of satellite DNAs of Mustelidae].

DNA reassociation kinetics were studied in the European mink (Mustela lutreola), the American mink (M. vison), the marbled polecat (Vormela peregusna). Variation in DNA quantity and heterochromatin amount occurs in connection with changes in the size of all kinetic fractions. Moderately repetitive genome component is the most variable in these three species. Cryptic CsCl satellite of the stoat (M. erminea), Ag+/Cs2SO4 satellites of the M. vison, V. peregusna were used for in situ homo- and heterologous hybridizations. Satellite DNAs revealed may be classified for the evolution age and chromosomal location type. More ancient satellite DNAs were dispersed in carnivors or mammalian genomes. Mustelids' specific satellites are concentrated in heterochromatic chromosome regions. The evolutionary implications of these findings are discussed.

Animals↗

[Homology of G-banded mammalian chromosomes].

Linkage relationships of homologous loci and high resolution G-banding patterns of man, mouse, rat, chinese hamster, rabbit, cat, mink, pig, ox and sheep were used for identification of 11 evolutionary conservative autosomal regions. The distributions and inversions of these regions in the ancestor genomes of some phylums have been shown. For example, the regions homologous to human Ip region were detected in cat, mink and rabbit genomes. In the genomes of rodents studied the intraregion inversion was detected. In the ox and sheep genomes the distal end deletions were detected within these regions. In the pig genome these regions were represented solely by disruptions. We supposed that the rapid "catastrophic" chromosomal evolution took place during short time periods of some orders and families separation.

Animals↗

[Characteristics of the distribution of DNA repetitive sequences in the sex chromosomes of 4 species of rodent].

Four rodent species with very large heterochromatic regions on the sex chromosomes have been studied using in situ DNA/DNA hybridization techniques. Repetitious DNA fractions were obtained at C0t 0-0.01. Heterochromatic regions of X and X chromosomes of Cricetulus barabensis and Phodopus sungorus, and the heterochromatic long arm of the Y chromosome of Mesocricetus auratus do not contain disproportionately high amounts of repeated DNA sequences. Heterochromatic regions on sex chromosomes of Microtus subarvalis contain high amounts of repeated DNA sequences. Additional heterochromatic autosomal arms, a heterochromatic arm of the X chromosome, and a short arm of the Y chromosome of Mesocricetus auratus contain high amounts of repeated DNA sequences too.

Animals↗

[Cytogenetics of albinism in polecats of the genus Putorius (Carnivora, Mustelidae)].

Karyotypes of two ferret species, Putorius eversmanni (2n = 38) and P. putorius (2n = 40), differ in a single Robertsonian rearrangement, resulting in decreased chromosome number in P. eversmanni. Interspecific hybrids from crosses between P. eversmanni and domesticated albino ferret P. putorius furo are fertile. Hybrid individuals have a chromosome set 2 = 39. Analysis of offspring from crosses of these hybrids showed that albinism is a recessive character. Offspring obtained from crosses between interspecific hybrid ferrets and between these hybrids and parental species was subjected to chromosome analysis. A statistically significant correlation was observed between the chromosome number and albinism. As a rule, albino hybrids have a chromosome number 2n = 40. This suggests that the locus, controlling albinism, is located at one of the two pairs of acrocentric chromsomes involved in Robertsonian rearrangement. The rare occurrence of albino hybrids with 2n = 39 may be attributed to recombination. According to our data, recombination probability is less than 0.5 which indicates that the albinism locus located near the centromere. Possible extrapolation of the data obtained for ferrets to other Mustelidae is discussed.

Albinism↗

[Development and certification of libraries of the MDCK continuous cell line for production of influenza vaccine].

Seeding and working banks of the continuous MDCK cell culture suitable for the production of cultured influenza vaccine were created and deposited at liquid nitrogen temperature at the "Vector" State Research Center of Virology and Biotechnology. The MDCK cell culture was shown to have morphology typical of the discussed cell line; it does not have any alien agents and is oncogenically safe; its enzimogram and karyotype are typical of the donor line; finally, its biological properties are stable during a long period of cultivation and its sensitivity to influenza virus is high, therefore, it can be recommended for the production of influenza vaccine. The continuous MDCK cell line was certified at Tarasevich Committee and was recommended by the MIBP Committee, Russia's Health Ministry, for its use as a substrate in the production of diagnostic and preventive immunoglobulins.

Animals↗

[Interstitial telomere repeats as markers of evolutionary changes in the mammalian karyotype: human chromosome 2].

Telomer repeats represented by hexamer (TTAGGG)n at chromosome termini are required for correct function and chromosome stability. At the same time, interstitial telomer sequence (ITS) located far from the chromosome ends are known for several mammalian genomes, including the human genome. It is assumed that these repeats mark the points of fusion or other chromosome reconstructions of ancestors. Exact localization of all interstitial telomer sequences in the genome could greatly improve our understanding of the mechanism of karyotype evolution and species origin. We have developed a software for a search of interstitial telomer sequences in complete sequences of mammalian genomes. We have demonstrated the evolutionary significance of repeats by an example of human chromosome 2. The results and supplementary materials are available at the site of the Institute of Cytology and Genetics: http://www.bionet.nsc.ru/labs/theorylabmain/orlov/telomere/.

Chromosomes, Human, Pair 2↗

[A new case of trisomy in cattle].

The new type of trisomy (2n = 61, XX, +19) was found in the heifer with prognathia inferior syndrome. Correlations between trisomy of different types and phenotypic abnormalities are discussed.

Animals↗

[A new chromosomal rearrangement in cattle].

The karyotype of a heifer with 2n-59 was studied using routine, C- and high-resolution G-banding techniques. A 1/29 translocation was identified. One of X chromosomes was normal. Another X chromosome was absent, but G-positive, partly heterochromatic biarmed element was present in all the cells. It is supposed that deletion of greater part of the q-arm of the second X chromosome and heterochromatinization of the remains took place.

Animals↗