[NotI-STS markers for human chromosome 3 are gene markers].

Ninety four NotI-STS markers to seventy two individual NotI clones were developed basing on DNA nucleotide sequences from NotI-"jumping" and "linking" NotI-libraries of human chromosome 3. The localization of NotI-STS markers and their ordering on chromosome was established by combined data of RH-mapping (our data), contig-mapping, cytogenetic mapping and in silico mapping. Performed comparison of NotI-STS DNAs with human genome sequences revealed two gaps in the regions, 3p21.33 (marker NLI-256) and 3p21.31 (NL3-005), and segmental duplication. Identical DNA fragments are localized in the regions 12q and 3p22-21.33 (marker NL3-007). In the region 3q28-q29 (marker NLM-084) a fragment was detected with its identical copies present also on chromosomes 1, 2, 15 and 19. For 69 NotI-STSs, significant homologies with nucleotide sequences of 70 genes and two cDNAs were detected taking in consideration homologies to NotI-STS 5'- and 3'-terminal sequences. Association of NotI-STSs with genes is confirmed by high correlation of gene density distribution with the density of NotI-STS markers on the map of human chromosome 3. Obtained data evidence possibility of NotI-STS marker application as gene markers and allow considering constructed NotI-map as gene map of human chromosome 3.

[The structure of the human oncogenesis-associated CKAP2 (LB1) gene]. / Struktura sviazannogo s onkogenezom gena CKAP2 (LB1) cheloveka.

The structure was described for human CKAP2, which codes for the cytoskeleton-associated protein 2, is at the boundary of chromosome regions 13q14.3 and 13q21.1, and is rearranged in various tumors. The CKAP2 exonintron structure was established by collating the nucleotide sequences of the cDNA and genomic clone AL359513 (GenBank) and analyzing the gene sequence with the GENSCAN program. The results were verified by amplifying gene fragments. CKAP2 comprises nine exons ranging 70-1442 bp and is about 22 kb in size (regulatory regions included). The CKAP2 promoter contains CCAAT (-39...-33) rather than the canonical TATA box, and harbors nine binding sites for six transcription factors. Thus, CKAP2 possesses all structural elements characteristic of eukaryotic genes. The results may be used to study the CKAP2 expression in normal and tumor cells in order to elucidate its role in carcinogenesis.

[Evolutionarily-conserved gene CKAP2,located in region 13q14.3 of the human genome, is frequently rearranged in various tumors]. / Evoliutsionno-konservativnyi gen CKAP2 raspolozhen v oblasti genoma cheloveka 13q14.3, chasto perestraivaemoi v razlichnykh opukholiakh.

The human CKAP2 gene, which is involved in diffuse large B-cell lymphomas, was localized via screening the GeneBridge 4 somatic cell radiation hybrid panel by means of the polymerase chain reaction (PCR). The CKAP2 gene was mapped between the WI-15460 and WI-3673 markers at the boundary between regions 13q14.3 and 13q21.1, at the distance of 14.39 cR (with 4.8 cR per cM) from the WI-5867 framework marker (lod score > 2.26). The human CKAP2 gene displayed high homology to mouse and rat expressed orthologs, A CKAP2-like sequence was found in human chromosome 14 and assumed to be a pseudogene resulting from duplication and subsequent mutations of the CKAP2 gene on chromosome 13. A possible role of the CKAP2 gene in oncogenesis associated with deletions and rearrangements of region 13q14.3-21.1 is discussed.

[Mapping of EST- and STS-markers in the human genome using a panel fo radiation hybrids]. / Kartirovanie v genome cheloveka EST- i STS-markerov s ispol'zovaniem paneli radiatsionnykh gibridov.

Ten DNA markers were localized in the human genome by a screening procedure against the radiation hybrid somatic cell panel (GeneBridge 4 RH Panel) using polymerase chain reaction (RH mapping method). DNA markers were developed to nucleotide sequences adjacent to NotI sites of human chromosome 3 (NotI-STS markers) and also to nucleotide sequences of human cDNA (EST markers). Three EST markers mapped (B10164, S16R and 18F5R) were localized in the human genome for the first time. Marker B10164 was found to be homologous to the nucleotide sequence of the BASP1 gene coding a major receptor protein. Markers S16R and 18F5R presumably tagged new genes, because no homologies were revealed among the nucleotide sequences presented in the databases. For four NotI-STS, more precise localization on human chromosome 3 was determined. On the basis of the data obtained, the NotI map may be integrated with other types of physical maps of human chromosome 3. RH mapping with a standard commercial panel of radiation hybrid somatic cells provided a chance to integrate the data obtained into international databases and existing integrated human chromosomal maps.

[Radiation mapping as one of main methods of creating high resolution maps of human and animal genomes]. / Radiatsionnoe kartirovanie kak odin iz osnovnykh metodov postroeniia fizicheskikh kart genomov cheloveka i zhivotnykh s vysokim urovnem razresheniia.

Radiation hybrid mapping (RH mapping) is considered as one of the main methods of constructing physical maps of mammalian genomes. In introduction, theoretical prerequisites of developing of the RH mapping and statistical methods of data analysis are discussed. Comparative characteristics of universal commercial panels of the radiation hybrid somatic cells (RH panels) are shown. In experimental part of the work, RH mapping is used to localise nucleotide sequences adjacent to NotI sites of human chromosome 3 with the aim to integrate contig map of NotI clones to comprehensive maps of human genome. Five nucleotide sequences adjacent to the sites of integration of papilloma virus in human genome and expressed in the cells of cervical cancer were localised. It was demonstrated that the region 13q14.3-q21.1 was enriched with nucleotide sequences involved in the processes of oncogenesis. RH mapping can be considered as one of the most perspective applications of the modern radiation biology in the field of molecular genetics, that is, in constructing physical maps of mammalian genomes with high resolution level.