Alu-PCR approach to isolating NotI-linking clones from the 3p14-p21 region frequently deleted in renal cell carcinoma

Journal Article · Tue Jun 01 04:00:00 EDT 1993 · Genomics; (United States)

DOI:https://doi.org/10.1006/geno.1993.1252· OSTI ID:7076701

Zabarovsky, E R ^[1]; Kashuba, V I ^[2]; Pokrovskaya, E S; Zabarovska, V I; Klein, G; Winberg, G ^[3]; Wang, Ji Yi; Berglund, P; Sumegi, J ^[4]; Boldog, F ^[5]

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1. Karolinska Institute, Stockholm (Sweden) Engelhardt Institute of Molecular Biology, Moscow (Russian Federation)
2. Karolinska Institute, Stockholm (Sweden) Institute of Molecular Biology and Genetics, Kiev (Ukraine)
3. Karolinska Institute, Stockholm (Sweden)
4. Univ. of Nebraska Medical Center, Omaha (United States)
5. Univ. of Colorado Health Science Center, Denver, CO (United States); and others

In the mammalian genome, CpG islands are associated with functional genes and cloning of these islands could be an alternative approach for cloning functional genes. Recently the authors have developed a new approach for cloning CpG islands and constructing NotI linking libraries. They have initiated the construction of a NotI restriction map for chromosome 3, especially focusing on the rearrangements in the 3p14-p21 region, which are associated with different malignancies. CpG islands from this region are useful for isolation of candidate tumor suppressor genes that map to this region and for isolating NotI-linking clones from 3p14-p21 for mapping purposes. Here they suggest a modification of Alu-PCR as an approach to isolating NotI sites (e.g., CpG islands) from defined regions of the chromosome. Instead of using whole chromosomal DNA for Alu-PCR, they have used representative NotI-linking libraries from hybrid cell lines containing either whole or deleted human chromosome 3 (MCH903.1 and MCH924.4, respectively). This decreases the complexity of the Alu-PCR products 1-100 times compared to the whole human genome. Using this modification, they can isolate NotI-linking clones, which are natural markers on the chromosome, rather than random genomic fragments. Among eight clones selected by this method, seven were from the region deleted in MCH924.4. The results clearly demonstrate the feasibility of Alu-PCR for isolating CpG islands from defined regions of the genome. 32 refs., 5 figs., 1 tab.

• OSTI ID: 7076701
• Journal Information: Genomics; (United States), Journal Name: Genomics; (United States) Vol. 16:3; ISSN GNMCEP; ISSN 0888-7543
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
BODY
CARCINOMAS
CHROMOSOMES
CLONING
DISEASES
DNA HYBRIDIZATION
DNA-CLONING
GENES
GENETIC MAPPING
HUMAN CHROMOSOME 3
HUMAN CHROMOSOMES
HYBRIDIZATION
KIDNEYS
MAPPING
NEOPLASMS
ORGANS