Abstract
Development of the methods permitting cloning of identical sequences between two sources of DNA can be very useful for many purposes, including isolation of disease genes. Here we describe a new method called CIS (cloning of identical sequences). A combination of digestion with MvnI, treatment with mung bean nuclease, UDG (uracil–DNA glycosylase) and PCR with 5′-methyl-dCTP and dUTP was used to isolate identical sequences between two micro-cell hybrid lines (MCH). In a control experiment, mouse MCH903.1 and MCH939.2 containing human chromosome 3 from different individuals, were compared using the CIS procedure. Only background fluorescence in-situ hybridization (FISH) was achieved. In another experiment, mouse MCH903.1, containing complete human chromosome 3, and rat MCH429.11, containing a part of human 3q from the same chromosome were compared. The experiment showed that the original MCH429.11 and the DNA purified using the CIS procedure had identical FISH patterns to human metaphase chromosomes, thus demonstrating the efficiency of CIS.
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Au KG, Welsh K, Modrich P (1992) Initiation of methyl-directed mismatch repair. J Biol Chem 267: 12142–12148.
Cheung VG, Nelson SF (1998) Genomic mismatch scanning identifies human genomic DNA shared by identical descent. Genomics 47: 1–6.
Cheung VG, Gregg JP, Gogolin-Ewens KJ et al. (1998) Linkagedisequilibrium mapping without genotyping. Nat Genet 18: 225–230.
Ellis LA, Taylor GR, Banks R, Baumberg S (1994) MutS binding protects heteroduplex DNA from exonuclease digestion in vitro: a simple method for detecting mutations. Nucl Acids Res 22: 2710–2711.
Fedorova L, Kost-Alimova M, Gizatullin RZ et al. (1997) Assignment and ordering of twenty-three unique NotI-linking clones containing expressed genes including the guanosine 59-monophosphate synthetase gene to human chromosome 3. Eur J Hum Genet 5: 110–116.
Kholodnyuk ID, Kost-Alimova M, Kashuba VI et al. (1997) The region of 3p22-p21.3 is non-randomly eliminated from mouse human microcell hybrids during tumor growth in SCID mice. Genes Chromosomes Cancer 18: 200–211.
McAllister L, Penland L, Brown PO (1998) Enrichment for loci identical-by-descent between pairs of mouse or human genomes by genomic mismatch scanning. Genomics 47: 7–11.
Mirzayans F, Mears AJ, Guo SW, Pearce WG, Walter MA (1997) Identification of the human chromosomal region containing the iridogoniodysgenesis anomaly locus by genomic-mismatch scanning. Am J Hum Genet 61: 111–119.
Nelson SF, McCusker JH, Sander MA, Kee Y, Modrich P, Brown PO (1993) Genomic mismatch scanning: a new approach to genetic linkage mapping. Nat Genet 1: 11–18.
Pinkel D, Straume T, Gray JW (1986) Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc Natl Acad Sci USA 83: 2934–2938.
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd edn. New York: Cold Spring Harbor Press.
Wang JY, Zabarovsky ER, Talmadge C et al. (1994) Somatic cell hybrid panel and NotI linking clones for physical mapping of human chromosome 3. Genomics 20: 105–113.
Welsh KM, Lu AL, Clark S, Modrich P (1987) Isolation and characterization of the Escherichia coli mutH gene product. J Biol Chem 262: 15624–15629.
Zabarovsky ER, Kashuba VI, Pokrovskaya ES et al. (1993) Alu-PCR approach to isolate NotI linking clones from the 3p14–3p21 region frequently deleted in renal cell carcinoma. Genomics 16: 713–719.
Zabarovsky ER, Kashuba VI, Kholodnyuk ID et al. (1994) Rapid mapping of NotI linking clones with differential hybridization and Alu-PCR. Genomics 21: 486–489.
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Zabarovska, V., Li, J., Muravenko, O. et al. CIS – Cloning of Identical Sequences Between Two Complex Genomes. Chromosome Res 8, 77–84 (2000). https://doi.org/10.1023/A:1009243606611
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DOI: https://doi.org/10.1023/A:1009243606611