| First Author | Elliott RW | Year | 1992 |
| Journal | Mouse Genome | Volume | 90 |
| Issue | 3 | Pages | 428-29 |
| Mgi Jnum | J:2503 | Mgi Id | MGI:51026 |
| Citation | Elliott RW, et al. (1992) High resolution blot analysis of alleles for Glut-1 in RI strains. Mouse Genome 90(3):428-29 |
| abstractText | Full text of Mouse Genome contribution: HIGH RESOLUTION BLOT ANALYSIS OF ALLELES FOR GLUT-1 IN RI STRAINS. Rosemary W. Elliott, Joanne Pazik and Colleen Hohman. Department of Molecular and Cellular Biology, Roswell Park Cancer Inst, Elm and Carton, Buffalo, NY, 14263 Introduction. The genetic locus for the Glut-1 (glucose transporter-1) has been shown to be on Chr 4 in the interval between Ifa (interferon alpha) and Lck (lymphocyte protein tyrosine kinase) by Bahary et al (1), who used a backcross involving C57BL/6J and M. spretus. We have employed high resolution blot analysis (3) to find a DNA variant between inbred strains, and used this to determine the strain distribution pattern in the BXD, AKXD and BXH RI strains. Materials and Methods. Plasmid pGT25L (10) was the kind gift of Dr. M. Mueckler of the Whitehead Inst. All methods including high resolution Southern blotting of sequencing gels containing HaeIII-digested mouse DNA were performed as in Elliott et al. (3). Results. A search among inbred strains for DNA fragment size differences using twelve endonucleases with 6-base recognition sites and four endonucleases with 4-base recognition sequences was unsuccessful. However, use of HaeIII, followed by fragment separation in a sequencing gel produced 12 fragments in the size range 80 to 280 bp. In DNA from C57BL/6J, the variant fragment is 202 bp (allele Glutb), while in DNA from DBA/2J, the variant fragment is 200 bp (allele Glutd). Other strains carrying the Glutb allele are AKR/J, C57L/J, C58/J, NZB/BINJ, SM/J and SWR/J, while strains A/J, BALB/cByJ and C3H/HeJ carry the Glutd allele. Analysis of segregation of these fragments among the BXD, AKXD and BXH RI strains produced the strain distribution patterns (SDPs) shown in Table 1. The BXD SDP is identical to those reported for Mtv13 (7; 12), Cyp4a (9) and Ms15-1 (6) while in the AKXDs there are recombinants with Mtv-13 in strains 3, 8 and 28. The order of loci whose alleles were typed in at least 2 RI sets is Ifa -- Pmv-23 -- Glut-1 -- Ck-1 -- Lck. Distance estimates are shown at the end of Table 1. Discussion. These results illustrate the power of high resolution blot analysis to find DNA variation between inbred strains in cases where endonucleases with 6-base recognition sites revealed no differences. The combination of sequencing gels and an endonuclease with a 4-base recognition site allowed the detection of a fragment size differences of only 2 base pairs. Genetic analysis has been restricted to RI strains, but the locus may also be typed in appropriate backcrosses. Finding a fragment size difference between inbred strains has extended the usefulness of the Glut-1 locus as a genetic marker. References 1. Bahary, N., Zorich, G., Pachter, J.E., Leibel, R.L. and Friedman, J.M. 1991. Molecular genetic linkage maps of mouse Chromosomes 4 and 6. Genomics 11: 33-47. 2. Cho, M., Villani, V. and D'Eustachio, P. 1991. A linkage map of distal mouse chromosome 12. Mammalian Genome 1:30-36. 3. Elliott, R.W., Sparkes, R.S., Mohandas, T., Grant, S.G. and McGinnis, J.F. 1990. Localization of the Rhodopsin gene to the distal half of mouse chromosome 6. Genomics 6: 635-644. 4. Frankel, W.N., Stoye, J.P., Taylor, B.A. and Coffin, J.M. 1989. Genetic identification of endogenous polytropic proviruses by using recombinant mice. 5. Huppi, K., Mock, B.A., Schricker, P., D'Hoostelaere, L.A. and Potter, M. 1988. Organization of the distal end of mouse chromosome 4. Current Topics in Microbiol. and Immunol. 137: 276-288. 6. Jeffreys, A.J., Wilson, V., Kelly, R., Taylor, B.A. and Bullfield, G. 1987. Mouse DNA fingerprints: analysis of chromosomal localization and germ-line stability of hypervariable loci in recombinant inbred strains. Nucl. Acid Res. 15: 2823-2836. 7. Lee, B.K. and Eicher, E.M. 1990. Segregation patterns of endogenous mouse mammary tumor viruses in five recombinant Inbred strain sets. J. Vlrol. 64: 4568-4572. 8. Manly, K.F. and Elliott, R.W. 1991. Rl Manager, a microcomputer program for analysis of data from recombinant inbred strains. Mammalian Genome 1: 123-126. 9. Miles, J.S., Moss, J.E., Taylor, B.A., Burchell, B. and Wolf, C.R. 1991. Mapping genes encoding drug-metabolizing enzymes in recombinant inbred mice. Genomics 11: 309-316. 10. Mueckler, M., Caruso, C., Baldwin, S.A., Panico, M., Blench, I., Morris, H.R., Allard, W.J., Lienhard, G.E. and Lodish, H.F. 1985. Sequence and structure of a human transporter gene. Science 229: 941-945. 11. Nadeau, J.H., Berger, F.G., Kelley, K.A., Pitha, P.M., Sidman, C.L. and Worrall, N. 1986. Rearrangement of genes located on homologous chromosomal segments in mouse and man: the location of genes for alpha- and beta-interferon, alpha-1 acid glycoprotein-1 and Ð2, and aminolevulinate dehydratase on mouse chromosome 4. Genetics 104: 1239-1255. 12. Traina, V.L., Taylor, B.A., and Cohen, J.C. Genetic mapping of endogenous murine mammary tumor viruses: Locus characterization, segregation and chromosomal distribution. 13. Siracusa, L.D., Buchberg, A.M., Copeland, N.G. and Jenkins. 1989. Recombinant inbred-strain and interspecific backcross analysis of the molecular markers flanking the murine agouti coat color locus. Genetics 122: 669-679. Table 1. (Legend). SDP of GT25L-hybridizing HaeIII-fragments in RI strains. BXD Strain: Ifa: 1:D; 2: B; 5: B (X); 6: B; 8: B; 9: D; 11: B; 12: D; 13: D (X); 14: B (X); 15: D; 16: B (X); 18: B (X); 19: D; 20: B; 21: D; 22: D; 23: B (X); 24: D (X); 25: D; 27: D; 28: D; 29: B (X); 30: D; 31: B; 32: B (X). Glut: 1: D; 2: B; 5: D; 6: B; 8: B (X); 9: D; 11: B; 12: D; 13: B (X); 14: D; 15: D; 16: D; 18: D; 19: D; 20: B; 21: D; 22: D (X); 23: D; 24: B; 25: D; 27: D; 28: D; 29: D; 30: D; 31: B; 32: D. Ck-1: 1: D; 2: B; 5: D (X); 6: B; 8: D; 9: D; 11: B; 12: D; 13: D; 14: D; 15: D (X); 16: D; 18: D; 19: D; 20: B; 21: D; 22: B; 23: D; 24: B; 25: D; 27: D (X); 28: D; 29: D (X); 30: D; 31: B; 32: D. Lck: 1: D; 2: B; 5: B; 6: B; 8: D; 9: D; 11: B; 12: D; 13: D; 14: D; 15: B; 16: D; 18: U; 19: D; 20: B; 21: D; 22: B; 23: D; 24: B; 25: D; 27: B; 28: D; 29: B; 30: D; 31: B; 32: D. AKXD Strain: Pmv-23: 1: D; 2: D; 3: A (X); 6: D; 7: D; 8: A; 9: D; 10: A; 11: D; 12: A; 13: D; 15: D; 16: D; 18: D; 20: D; 21: A; 22: D (X); 23: A; 24: D; 25: D; 26: D; 27: D; 28: A (X). Glut: 1: D; 2: D; 3: D; 6: D; 7: D (X); 8: A (X); 9: D; 10: A; 11: D; 12: A; 13: D; 15: D; 16: D; 18: D; 20: D; 21: A; 22: A; 23: A; 24: D; 25: D; 26: D; 27: D; 28: D. Ck-1: 1: D; 2: D; 3: D; 6: D; 7: A; 8: D; 9: D; 10: A; 11: D; 12: A; 13: D; 15: D; 16: D; 18: D; 20: D; 21: A; 22: A; 23: A; 24: D; 25: D; 26: D; 27: D; 28: D. BXH Strain: Pmv-23/Ifa: 2: H; 3: B; 4: H; 6: B (X); 7: B; 8: B; 9: H; 10: B; 11: H; 12: B; 14: B; 19: H. Glut-1: 2: H; 3: B; 4: H; 6: H (X); 7: B (X); 8: B; 9: H; 10: B; 11: H; 12: B; 14: B (X); 19: H. Ck-1: 2: H (X); 3: B; 4: H; 6: B; 7: H; 8: B; 9: H; 10: B; 11: H; 12: B; 14: H; 19: H. Lck: 2: B; 3: B; 4: H; 6: B; 7: H; 8: B; 9: H; 10: B; 11: H; 12: B; 14: H; 19: H. Interval: Ifa Ð Glut-1; R: 10; N: 38; % rec. +/- S.D.: 10.9 +/- 4.9; 95% conf. limits: 4.2 Ð 30.5; 99% conf. limits: 3.1 Ð 43.3. Interval: Pmv-23 Ð Glut-1; R: 4; N: 35; % rec. +/- S.D.: 3.5 +/- 2.0; 95% conf. limits: 0.8 Ð 1. 1.2; 99% conf. limits: 0.5 Ð 15.2. Interval: Glut-1 Ð Ck-1; R: 8 (correction); N: 61; % rec. +/- S.D.: 3.5 +/- 1.5; 95% conf. limits: 1.3 Ð 8.3; 99% conf. limits: 0.9 Ð 10.5. Interval: Glut-1 Ð Lck; R: 11 (correction); N: 37; % rec. +/- S.D.: 11.4 +/- 5.2; 95% conf. limits: 4.3 Ð 32.6; 99% conf. limits: 3.2 Ð 47.1. A, B, D and H are generic symbols for alleles inherited from strains AKR/J, C57BL/6J, DBA/2J and C3H/HeJ respectively. A U indicates that the allele is nto known. SDPs for Ck-1 (2), Ifa (11), Lck (5; 13) and Pmv-3 (4) have been published. Calculations were performed using RI Manager ver. 2.2 (8). |
