Molecular mapping of the human major histocompatibility complex by pulsed-field gel electrophoresis

Journal Article · Thu Oct 01 00:00:00 EDT 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.20.7237· OSTI ID:6939401

Dunham, I; Sargent, C A; Trowsdale, J; Campbell, R D

Pulsed-field gel electrophoresis and cosmid walking have been used to establish a molecular map of the human major histocompatibility complex (MHC). The authors have isolated approx. = 230 kilobases (kb) of genomic DNA in overlapping cosmid clones covering the genes for the second and fourth components of complement (C2 and C4, respectively), factor B, and steroid 21-hydroxylase, and approx. = 82 kb of genomic DNA surrounding the genes for the tumor necrosis factors ..cap alpha.. and ..beta... Single-copy hybridization probes isolated from these cosmid clusters and probes for the known MHC gene loci were hybridized to Southern blots of genomic DNA that had been digested with infrequently cutting restriction endonucleases and separated on pulsed-field gels. The data obtained allowed the construction of a long-range genomic restriction map and indicated that the MHC spans 3800 kb. This map orients the MHC class III gene cluster with respect to the DR subregion; the C2 gene is on the telomeric side of the 21-hydroxylase B gene. In addition they have defined the positions of the genes for the tumor necrosis factors ..cap alpha.. and ..beta.. in the human MHC. Genes for the ..cap alpha.. chain of DR and 21-hydroxylase B are separated by at least 300 kb, while the distance between the genes for C2 and tumor necrosis factor ..cap alpha.. is 390 kb. The HLA-B locus lies approx. = 250 kb on the telomeric side of the tumor necrosis factor genes.

• Research Organization: Medical Research Council Immunochemistry Unit, Oxford (England)
• OSTI ID: 6939401
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 84:20; ISSN PNASA
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
62 RADIOLOGY AND NUCLEAR MEDICINE
AUTORADIOGRAPHY
CARBOHYDRATES
DISEASES
DNA
ELECTROPHORESIS
ENZYMES
GENES
GENETIC MAPPING
GLUCOPROTEINS
HYBRIDIZATION
HYDROXYLASES
IMMUNITY
MAPPING
NECROSIS
NEOPLASMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PATHOLOGICAL CHANGES
PROTEINS
SACCHARIDES