Mapping of HLA genes using pulsed‐field gradient electrophoresis

Ragoussis, Jiannis; Bliek, Alexander van der; Trowsdale, John; Ziegler, Andreas

doi:10.1016/0014-5793(86)81376-x

Cited by 21 publications

(15 citation statements)

References 13 publications

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“…This technique has been used to determine the arrangement of the class II genes within a region of 900 kb (14) and to provide a version of the whole MHC (15) slightly different from that presented here and not including the TNF genes. In addition, the class II and class III regions were linked within a common 1000-kb Not I restriction fragment (15,16). These studies suggested, on the basis of a linear alignment of restriction fragments hybridizing with class II, class III, or class I coding sequence probes, that the entire MHC spans about 2000-3000 kb.…”

mentioning

confidence: 91%

Linkage map of the human major histocompatibility complex including the tumor necrosis factor genes.

Carroll

Katzman

Alicot

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

297

136

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Linkage map of the human major histocompatibility complex including the tumor necrosis factor genes (pulsed- ABSTRACTThe tumor necrosis factor (TNF) a and (3 gene pair has been linked in the human major histocompatibility complex to HLA-B, HLA-C, and, tentatively, HLA-E and HLA-A on one side and to the class III complement/steroid, 21-hydroxylase gene cluster on the other by pulsed-field gel electrophoresis. The TNF genes are located 200 kilobases (kb) centromeric of HLA-B and about 350 kb telomeric of the class III cluster. Together with previous data on the linkage and structures of the class II and class III regions, a restriction map of the entire human major histocompatibility complex of about 3500 kb has been prepared.

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mentioning

confidence: 91%

Linkage map of the human major histocompatibility complex including the tumor necrosis factor genes.

Carroll

Katzman

Alicot

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

297

136

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“…Using PFGE, Hardy et al (10) determined a map for the human class II region and established the order of the HLA-D subregions. Physical linkage between the human class II and class III loci has also been demonstrated using this technique (11,12). In mouse the orientation and molecular map position of the complement gene cluster have been established (13).…”

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confidence: 99%

“…The technique of pulsed-field gel electrophoresis (PFGE) (8,9), which allows the resolution of DNA fragments >2 megabases, in combination with restriction enzymes that cut rarely in the mammalian genome, has been used for long-range restriction-site mapping of the MHC in man (10)(11)(12) and in mouse (13). Using PFGE, Hardy et al (10) determined a map for the human class II region and established the order of the HLA-D subregions.…”

mentioning

confidence: 99%

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

Dunham

Sargent

Trowsdale

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

211

107

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Pulsed-field gel electrophoresis and "cosmid walking" have been used to establish a molecular map of the human major histocompatibility complex (MIC). We have isolated -230 kilobases (kb) of genomic DNA in overlapping cosmid clones covering the genes for the second and fourth components ofcomplement (C2 and C4, respectively), factor B, and steroid 21-hydroxylase, and --=82 kb of genomic DNA surrounding the genes for the tumor necrosis factors a and (. 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 we have defined the positions of the genes for the tumor necrosis factors a and .8 in the human MHC. Genes for the a 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 a is 390 kb. The HLA-B locus lies -250 kb on the telomeric side of the tumor necrosis factor genes.The human major histocompatibility complex (MHC) is located on the short arm ofchromosome 6 in the distal portion of the 6p21.3 band (1). It consists of three major linked gene clusters. The class I and class II regions each encode highly polymorphic families of cell-surface glycoproteins involved in immune regulation. The class I loci consist of at least 17 highly related genes (2) that include those encoding the classical transplantation antigens (HLA-A, -B, and -C). The class II region (HLA-D) is arranged into four subregions DP, DZ/DO, DQ, and DR, each containing at least one a-and ,8-chain pair of genes (3). The class III region contains the genes encoding the serum complement proteins factor B and the second and fourth components of complement (C2 and C4, respectively), as well as two copies of the steroid 21-hydroxylase (21-OHase) genes that are closely linked to the two C4 loci, C4A and C4B (4, 5). Analysis of recombinant HLA haplotypes in family studies has established that the class I loci are telomeric to the class II genes. Within the class II region the DP subregion maps on the centromeric side of DQ and DR. Located between the HLA-DR and HLA-B loci are the class III genes, but the orientation and distance of these genes relative to the class I and class II genes have not been determined. The possibility that a number of other genes may reside within the MHC was exemplified by the finding that the genes for the tumor necrosis factors a and 3 (TNFa and TNFf, respectively) are linked to the HLA genes (6) and map close to the H-2D region in mouse (7).Clusters of overlapping cosmid clones have been isolated from the subregions of the class II loci (3) and fro...

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“…The physical length of the entire HLA complex is unknown so far, but our estimate based on the separation of DNA fragments containing HLA genes by pulsed-field gel electrophoresis (Schwartz andCantor 1984, Carle andOlson 1984) indicates that it encompasses at least 2500 kb pairs (Ragoussis et al 1986). This estimate has recently been confirmed (Lawrance et al 1987).…”

mentioning

confidence: 96%

“…Mutant BM 19.7 is a monosomy 6 mutant cell line retaining the A2 haplotype (Ziegler et al 1985b). BM 28.7 also exhibits monosomy 6, but with loss of the chromosome bearing the A2 haplotype (Ragoussis et al 1986). In the interstitial deletion mutant BM 2.2.3, the class I region of the A2 haplotype has been deleted, but the class II and III regions of this haplotype are still present, as is the chromosome 6 carrying the A1 haplotype (Ziegler et al 1985a).…”

mentioning

confidence: 99%

Localization of the genes for tumor necrosis factor and lymphotoxin between the HLA class I and III regions by field inversion gel electrophoresis

Ragoussis

Bloemer²,

Weiß³

et al. 1988

Immunogenetics

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The human major histocompatibility (HLA) complex is located on the short arm of chromosome 6 in the 6p21. 31-6p21.33 region (Spring et al. 1985, Ziegler et al. 1985a). The physical length of the entire HLA complex is unknown so far, but our estimate based on the separation of DNA fragments containing HLA genes by pulsed-field gel electrophoresis (Schwartz and Cantor 1984, Carle andOlson 1984) indicates that it encompasses at least 2500 kb pairs (Ragoussis et al. 1986). This estimate has recently been confirmed (Lawrance et al. 1987). Apart from the highly polymorphic class I and II loci, the HLA complex contains the genes for several complement components and 21-steroid hydroxylase (class III region) (Lamm and Olaisen 1985). In addition, the loci for tumor necrosis factor (TNFA) as well as lymphotoxin (TNFB) are also near or even within the HLA region (Nedwin et al. 1985). Spies and co-workers (1986) demonstrated that the TNF genes map either centromeric to HLA-DP or telomeric to the class I1 region, although apparently not in the vicinity of any known class I or III genes. The recent demonstration that the TNF loci are situated 70 kb upstream of the H-2D gene in the BALB/c mouse between the class III and class I regions (Mfiller et al. 1987a, b) suggested an analogous location in man, because the genetic organization of the major histocompatibility complexes (MHC) of both species is very similar. To clarify the position of the TNFA and TNFB genes on the HLA map, we have assigned TNFA to large DNA restriction fragments separated by field inversion gel electrophoresis (FIGE) (Carle et al. 1986), which hybridize with either class 1II-or class Ispecific probes as well. These results prove that the TNFA locus is localized between the HLA class III region and the HLA-B locus.To avoid interpretative difficulties which might arise from haplotype-specific restriction fragment length polymorphisms, mutant human cell lines with monosomy 6 or HLA hemizygosity were employed. All mutants were derived from BJAB-B95.8.6 lymphoma cells with the HLA haplotypes A1, Cw4,B35 and A2, C-,B13 (Spring et al. 1985). Mutant BM 19.7 is a monosomy 6 mutant cell line retaining the A2 haplotype (Ziegler et al. 1985b). BM 28.7 also exhibits monosomy 6, but with loss of the chromosome bearing the A2 haplotype (Ragoussis et al. 1986). In the interstitial deletion mutant BM 2.2.3, the class I region of the A2 haplotype has been deleted, but the class II and III regions of this haplotype are still present, as is the chromosome 6 carrying the A1 haplotype (Ziegler et al. 1985a).Large genomic DNA fragments were generated with various restriction enzymes, separated by FIGE, and analyzed by hybridization to Southern blots (Southern 1975) under stringent conditions. In the Bss HII digests (Fig. 1), a 370 kb DNA fragment was detected which hybridized both to the HLA-B locus-specific probe that had been derived 29 kb 5' of the HLA-B gene (Fig. lb) and to the TNFA probe (Fig. lc), indicating linkage of TNFA with the HLA-B locus. The class III loci ...

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Mapping of HLA genes using pulsed‐field gradient electrophoresis

Cited by 21 publications

References 13 publications

Linkage map of the human major histocompatibility complex including the tumor necrosis factor genes.

Linkage map of the human major histocompatibility complex including the tumor necrosis factor genes.

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

Localization of the genes for tumor necrosis factor and lymphotoxin between the HLA class I and III regions by field inversion gel electrophoresis

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