Abstract
We have explored the potential of irradiation-fusion gene transfer (IFGT) hybrids as a source of well-defined human chromosome fragments from which probes can be derived. Extensive characterization of the IFGT hybrid 4J4 with a full panel of markers from Chromosome (Chr) 6 showed that the human DNA content derives largely from 6p21.3 and 6q27. A cosmid library has been constructed from 4J4 DNA, and 370 recombinants containing human DNA have been isolated and overlapping clones ordered into 20 contigs. Regional localization of representative clones from each contig, determined by fluorescent in situ hybridization (FISH), places 13 contigs in 6q27 and 6 in 6p21.3. Preliminary screening of cDNA libraries with selected cosmids has identified two expressed sequences. Since there are a number of medically important genes in both these regions of human Chr 6 with several disease loci linked to the HLA-A region in 6p21.3 and various tumor suppressor genes to 6q27, this library will provide a valuable resource to aid the isolation of candidate genes for these diseases. In addition, unique markers for detailed physical and genetic mapping of these regions of human Chr 6 can be easily obtained.
Similar content being viewed by others
References
Adams, M.D., Dubnick, M., Kerlavage, A.R., Moreno, R., Kelley, J.M., Utterback, T.R., Nagle, J.W., Fields, C., Venter, J.C. (1992). Sequence identification of 2,375 human brain genes. Nature 355, 632–634.
Barbosa, J.A., Kamarck, M.E., Biro, A., Weisman, S.M., Ruddle, F.H. (1982). Identification of human genomic clones coding the major histocompatibility antigens HLA-A2 and HLA-B7 by DNA-mediated gene transfer. Proc. Natl. Acad. Sci. USA 79, 6327–6331.
Cox, D.R. (1992). Radiation hybrid mapping. Cytogenet. Cell Genet. 59, 80–81.
El Kahloun, A., Chauvel, B., Mauvieux, V., Dorval, I., Jouanolle, A., Gicquel, I., Le Gall, J., David, V. (1993). Localisation of seven new genes around the HLA-A locus. Hum. Mol. Genet. 2, 55–60.
Gaidano, G., Haupstein, R.S., Parsa, N.Z., Offit, K., Rao, P.H., Lenoir, G., Knowles, D.M., Chaganti, R.S.K., Dalla-Favera, R. (1992). Deletions involving two distinct regions of 6q in B-cell non-Hodgkin lymphoma. Blood 80, 1781–1787.
Gaidano, G., Shortle, V., Hauptschein, R., Venkatraji, V., Rutner, T., Dalla-Favera, R., Edwards, Y. (1993). HindIII, SacI and SSPI RFLPs at the D6S347 locus on a region of 6q which is frequently deleted in B-cell non-Hodgkin lymphoma and renal cell carcinoma (6q27). Hum. Mol. Genet., in press.
Glaser, T., Rose, E., Morse, H., Housman, D., Jones, A. (1990). A panel of irradiation hybrids selectively containing human chromosome 11p13: their structure and use to purify the WAGR gene complex. Genomics 6, 48–64.
Heinikoff, S. (1987). Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol. 155, 156.
Koller, B.H., Sidwell, B., Demars, R., Orr, H.T. (1984). Isolation of HLA locus specific DNA probes from the 3′ untranslated region. Proc. Natl. Acad. Sci. USA 81, 5175–5178
Kravitz, K., Skolnick, M., Canning, S., Carmelli, D., Baty, B., Amos, B., Johnson, A., Mendell, N., Edwards, C., Cartwright, G. (1979). Genetic linkage between hereditary haemochromatosis and HLA. Am. J. Hum. Genet. 31, 601–610.
Lichter, P., Chieh-Ju, C.T., Call, K., Hermannson, G., Evans, G.A., Housman, D., Ward, D.C. (1990). High resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones. Science 247, 64–69
Meese, E., Witkowski, C., Zohgbi, H., Stanbridge, E., Metzler, P., Trent, J. (1992). Development and utilisation of a somatic cell hybrid mapping panel to assign NotI linking clones to the long arm of human chromosome 6. Genomics 12, 542–548.
Ono, M., Yasunaga, T., Miyata, T., Ushikubo, H. (1986). Nucleotide sequence of human endogenous retrovirus genome related to the mouse mammary tumor virus genome. J. Virol. 60, 589–598.
Ragoussis, J., Jones, T.A., Sheer, D., Shrimpton, A.E., Goodfellow, P.N., Trowsdale, J., Ziegler, A. (1991). Isolation of probes specific to human chromosomal region 6p21 from immunoselected irradiation-fusion-gene transfer hybrids. Genomics 10, 598–607.
Saito, S., Saito, H., Koi, S., Sagae, S., Kudo, R., Saito, J., Noda, K., Nakamura, Y. (1992). Fine scale deletion mapping of the distal long arm of chromosome 6 in 70 human ovarian cancers. Cancer Res. 52, 5815–5817.
Sanger, F., Nicklen, S., Coulson, A.R. (1977). DNA sequencing with chain terminating inhibitors. Proc. Natl. Acad. Sci. USA 74, 5463.
Shortle, V., Malaspina, P., Dooley, J., Edwards, Y. (1993). Two novel cDNAs homologous to sequences on human chromosome 6. In preparation.
Simon, M., Bourell, M., Genetet, B., Fauchet, R. (1977). Idiopathic hemochromatosis: demonstration of recessive transmission and early detection by family HLA typing. N. Engl. J. Med. 297, 1017–1021.
Strachan, T., Sodoyer, R., Damotte, M., Jordan, B.R. (1984). Complete nucleotide sequence of a functional class I HLA gene, HLA-A3: implications for the evolution of HLA genes. EMBO J. 3, 887–894.
Sulston, J., Mallet, F., Staden, R., Durbin, R., Horsnell, T., Coulson, A. (1988). Software for genome mapping by fingerprinting techniques. Comput. Appl. Biosci. 4, 125–132.
Trent J., Ziegler, A. (1993). Report of the first international workshop on human chromosome 6 mapping 1992. Cytogenet. Cell Genet. 62, 65–87.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Shortle, V.P., Malaspina, P., Fox, M. et al. A cosmid library specific for human Chromosome regions 6p21.3 and 6q27. Mammalian Genome 4, 493–498 (1993). https://doi.org/10.1007/BF00364783
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00364783