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. 1987 Jun 11;15(11):4437–4448. doi: 10.1093/nar/15.11.4437

Cosmid mapping of the human chorionic gonadotropin beta subunit genes by field-inversion gel electrophoresis.

M Y Graham, T Otani, I Boime, M V Olson, G F Carle, D D Chaplin
PMCID: PMC340872  PMID: 3035494

Abstract

A cosmid clone containing the entire hCG beta gene cluster has been isolated. The restriction map of this clone has been determined by an indirect-end-label FIGE (field inversion gel electrophoresis) method. Analysis of this cosmid clone shows that there are 6 hCG beta genes in human genomic DNA. A previously uncloned portion of the hCG beta cluster, termed the "gap" region, has been shown not to contain any sequences homologous to the hCG beta cDNA. The restriction mapping method employed in this study takes advantage of the superior resolution of FIGE for high molecular weight DNA fragments in the size range 15-50 kb. This method is broadly applicable and permits rapid and accurate restriction mapping for extended regions of genomic DNA that have been cloned into cosmid or lambda vectors.

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Selected References

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  1. Boorstein W. R., Vamvakopoulos N. C., Fiddes J. C. Human chorionic gonadotropin beta-subunit is encoded by at least eight genes arranged in tandem and inverted pairs. Nature. 1982 Dec 2;300(5891):419–422. doi: 10.1038/300419a0. [DOI] [PubMed] [Google Scholar]
  2. Boothby M., Ruddon R. W., Anderson C., McWilliams D., Boime I. A single gonadotropin alpha-subunit gene in normal tissue and tumor-derived cell lines. J Biol Chem. 1981 May 25;256(10):5121–5127. [PubMed] [Google Scholar]
  3. Carle G. F., Frank M., Olson M. V. Electrophoretic separations of large DNA molecules by periodic inversion of the electric field. Science. 1986 Apr 4;232(4746):65–68. doi: 10.1126/science.3952500. [DOI] [PubMed] [Google Scholar]
  4. Carle G. F., Olson M. V. Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresis. Nucleic Acids Res. 1984 Jul 25;12(14):5647–5664. doi: 10.1093/nar/12.14.5647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chaplin D. D., Woods D. E., Whitehead A. S., Goldberger G., Colten H. R., Seidman J. G. Molecular map of the murine S region. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6947–6951. doi: 10.1073/pnas.80.22.6947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chu G., Vollrath D., Davis R. W. Separation of large DNA molecules by contour-clamped homogeneous electric fields. Science. 1986 Dec 19;234(4783):1582–1585. doi: 10.1126/science.3538420. [DOI] [PubMed] [Google Scholar]
  7. Cross S. H., Little P. F. A cosmid vector for systematic chromosome walking. Gene. 1986;49(1):9–22. doi: 10.1016/0378-1119(86)90381-1. [DOI] [PubMed] [Google Scholar]
  8. Fiddes J. C., Goodman H. M. The gene encoding the common alpha subunit of the four human glycoprotein hormones. J Mol Appl Genet. 1981;1(1):3–18. [PubMed] [Google Scholar]
  9. Grosveld F. G., Lund T., Murray E. J., Mellor A. L., Dahl H. H., Flavell R. A. The construction of cosmid libraries which can be used to transform eukaryotic cells. Nucleic Acids Res. 1982 Nov 11;10(21):6715–6732. doi: 10.1093/nar/10.21.6715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hussa R. O., Story M. T., Pattillo R. A. Cyclic adenosine monophosphate stimulates secretion of human chorionic gonadotropin and estrogens by human trophoblast in vitro. J Clin Endocrinol Metab. 1974 Feb;38(2):338–340. doi: 10.1210/jcem-38-2-338. [DOI] [PubMed] [Google Scholar]
  11. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Little P. F., Cross S. H. A cosmid vector that facilitates restriction enzyme mapping. Proc Natl Acad Sci U S A. 1985 May;82(10):3159–3163. doi: 10.1073/pnas.82.10.3159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Morgan F. J., Birken S., Canfield R. E. The amino acid sequence of human chorionic gonadotropin. The alpha subunit and beta subunit. J Biol Chem. 1975 Jul 10;250(13):5247–5258. [PubMed] [Google Scholar]
  14. Pierce J. G., Parsons T. F. Glycoprotein hormones: structure and function. Annu Rev Biochem. 1981;50:465–495. doi: 10.1146/annurev.bi.50.070181.002341. [DOI] [PubMed] [Google Scholar]
  15. Policastro P. F., Daniels-McQueen S., Carle G., Boime I. A map of the hCG beta-LH beta gene cluster. J Biol Chem. 1986 May 5;261(13):5907–5916. [PubMed] [Google Scholar]
  16. Policastro P., Ovitt C. E., Hoshina M., Fukuoka H., Boothby M. R., Boime I. The beta subunit of human chorionic gonadotropin is encoded by multiple genes. J Biol Chem. 1983 Oct 10;258(19):11492–11499. [PubMed] [Google Scholar]
  17. Rackwitz H. R., Zehetner G., Frischauf A. M., Lehrach H. Rapid restriction mapping of DNA cloned in lambda phage vectors. Gene. 1984 Oct;30(1-3):195–200. doi: 10.1016/0378-1119(84)90120-3. [DOI] [PubMed] [Google Scholar]
  18. Rackwitz H. R., Zehetner G., Murialdo H., Delius H., Chai J. H., Poustka A., Frischauf A., Lehrach H. Analysis of cosmids using linearization by phage lambda terminase. Gene. 1985;40(2-3):259–266. doi: 10.1016/0378-1119(85)90048-4. [DOI] [PubMed] [Google Scholar]
  19. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  20. Shome B., Parlow A. F. The primary structure of the hormone-specific, beta subunit of human pituitary luteinizing hormone (hLH). J Clin Endocrinol Metab. 1973 Mar;36(3):618–621. doi: 10.1210/jcem-36-3-618. [DOI] [PubMed] [Google Scholar]
  21. Smith H. O. Recovery of DNA from gels. Methods Enzymol. 1980;65(1):371–380. doi: 10.1016/s0076-6879(80)65048-4. [DOI] [PubMed] [Google Scholar]
  22. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  23. Talmadge K., Boorstein W. R., Vamvakopoulos N. C., Gething M. J., Fiddes J. C. Only three of the seven human chorionic gonadotropin beta subunit genes can be expressed in the placenta. Nucleic Acids Res. 1984 Nov 26;12(22):8415–8436. doi: 10.1093/nar/12.22.8415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Whittaker P. A., Southern E. M. Ultraviolet irradiation of DNA: a way of generating partial digests for rapid restriction site mapping. Gene. 1986;41(1):129–134. doi: 10.1016/0378-1119(86)90276-3. [DOI] [PubMed] [Google Scholar]

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