We have constructed a 12,000-rad porcine whole-genome radiation hybrid panel to complement the first generation 7,000-rad panel (IMpRH) and allow higher resolution mapping studies both in specific areas of interest and on the whole genome. We analyzed 243 hybrid clones on the basis of their marker retention frequency to produce a final panel of 90 hybrid clones with an average retention frequency of 35.4%. The resolution of this 12,000-rad panel (IMNpRH2) was compared to the resolution of the 7,000-rad panel (IMpRH) by constructing framework maps in the 2.4-Mb region of porcine chromosome 15 containing the acid meat RN gene. In this region, two-point analysis was used to estimate RH distances and demonstrates their reliability with the estimation of physical distances. This study demonstrates that the 12,000-rad panel constitutes a powerful tool for constructing high-resolution maps. Indeed, the resolution of IMNpRH2 (12–14 kb/cR12,000) is two to three times more than that of IMpRH (35–37 kb/cR7,000). As expected, the increase in the radiation dose allows an increase of the mapping resolution in terms of kb/cR with the same suppleness of use for mapping experiments. In addition the RH map constructed in the region investigated proved to be more homogeneous on IMNpRH2 than on IMpRH.   

1.
Alexander LJ, Troyer DL, Rohrer GA, Smith TP, Schook LB, Beattie CW: Physical assignments of 68 porcine cosmid and lambda clones containing polymorphic microsatellites. Mammal Genome 7:368–372 (1996).
2.
Boehnke M, Lange K, Cox DR: Statistical methods for multipoint radiation hybrid mapping. Am J hum Genet 49:1174–1188 (1991).
3.
Burton K: A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J 62:315–323 (1956).
4.
Demeure O, Renard C, Yerle M, Faraut T, Riquet J, Robic A, Schiex T, Rink A, Milan D: Rearranged gene order between pig and human in a QTL region on SSC 7. Mammal Genome in press.
5.
Gross-Belard M, Oudet P, Chambon P: Isolation of high molecular-weight DNA from mammalian cells. Eur J Biochem 36:32–38 (1973).
6.
Gyapay G, Schmitt K, Fizames C, Jones H, Vega-Czarny N, Spillett D, Muselet D, et al: A radiation hybrid map of the human genome. Hum molec Genet 5:339–346 (1996).
7.
Hawken RJ, Murtaugh J, Flickinger GH, Yerle M, Robic A, Milan D, Gellin J, et al: A first-generation porcine whole-genome radiation hybrid map. Mammal Genome 10:824–830 (1999).
8.
Jeon JT, Amarger V, Rogel-Gaillard C, Robic A, Bongcam-Rudloff E, Paul S, Looft C, et al: Comparative analysis of a BAC contig of the porcine RN region and the human transcript map: implications for the cloning of trait loci. Genomics 72:297–303 (2001).
9.
Jiang Z, Priat C, Galibert F: Traced Amplified sequence Tags (TOASTs) and mammalian comparative maps. Mammal Genome 9:577–587 (1998).
10.
Jones HB: Hybrid selection as a method of increasing mapping power for radiation hybrids. Genome Res 6:761–769 (1996).
11.
Lahbib-Mansais Y, Leroux S, Milan D, Yerle M, Robic A, Jiang Z, Andre C, et al: Comparative mapping between humans and pigs: localization of 58 anchorage markers (TOASTs) by use of porcine somatic cell and radiation hybrid panels. Mammal Genome 11:1098–1106 (2000).
12.
Lander ES, Linton LM, Birren B, Nusbaum C, Zody MC, Baldwin J, Devon K, et al: Initial sequencing and analysis of the human genome. Nature 409: 860–921 (2001).
13.
Lunetta KL, Boehnke M: Multipoint radiation hybrid mapping: comparison of methods, sample size requirements, and optimal study characteristics. Genomics 21:92–103 (1994).
14.
Lunetta KL, Boehnke M, Lange K, Cox DR: Selected locus and multiple panel models for radiation hybrid mapping. Am J hum Genet 59:717–725 (1996).
15.
McPherson JD, Marra M, Hillier L, Waterston RH, Chinwalla A, Wallis J, Sekhon M, et al: A physical map of the human genome. Nature 409:934–941 (2001).
16.
Milan D, Hawken R, Cabau C, Leroux S, Genet C, Lahbib Y, Tosser G, et al: IMpRH Server: an RH mapping server available on the Web. Bioinformatics 16:558–559 (2000a).
17.
Milan D, Jeon JT, Looft C, Amarger V, Robic A, Thelander M, Rogel-Gaillard C, et al: A mutation in PRKAG3 associated with excess glycogen content in pig skeletal muscle. Science 288:1248–1251 (2000b).
18.
Olivier M, Aggarwal A, Allen J, Almendras AA, Bajorek ES, Beasley EM, Brady SD, et al: A high-resolution radiation hybrid map of the human genome draft sequence. Science 291:1298–1302 (2001).
19.
Robic A, Seroude V, Jeon JT, Yerle M, Wasungu L, Andersson L, Gellin J, et al: A radiation hybrid map of the RN region in pigs demonstrates conserved gene order compared with the human and mouse genomes. Mammal Genome 10:565–568 (1999).
20.
Robic A, Jeon JT, Rey V, Amarger V, Chardon P, Looft C, Andersson L, et al: Construction of a high-resolution RH map of the human 2q35 region on TNG panel and comparison with a physical map of the porcine homologous region 15q25. Mammal Genome 12:380–386 (2001).
21.
Rohrer GA, Alexander LJ, Beattie CW: Mapping the beta subunit of follicle stimulating hormone (FSHB) in the porcine genome. Mammal Genome 5:315–317 (1994).
22.
Schiex T, Chabrier P, Bouchez M, Milan D: Boosting EM for radiation hybrid and genetic mapping. Proc of Workshop on Algorithms and Bioinformatics 2001, p 41–50 (2001).
23.
Schuler GD, Boguski MS, Stewart EA, Stein LD, Gyapay G, Rice K, White RE, et al: A gene map of the human genome. Science 274:540–546 (1996).
24.
Tosser-Klopp G, Bonnet A, Yerle M, Hatey F: Functional study and regional mapping of 44 hormono-regulated genes isolated from a porcine granulosa cell library. Genet Sel Evol 33:69–87 (2001).
25.
Vignaux F, Hitte C, Priat C, Chuat JC, Andre C, Galibert F: Construction and optimization of a dog whole-genome radiation hybrid panel. Mammal Genome 10:888–894 (1999).
26.
Yerle M, Pinton P, Robic A, Alfonso A, Palvadeau Y, Delcros C, Hawken R, et al: Construction of a whole-genome radiation hybrid panel for high-resolution gene mapping in pigs. Cytogenet Cell Genet 82:182–188 (1998).
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.