| First Author | Kelly CD | Year | 1993 |
| Journal | Mouse Genome | Volume | 91 |
| Issue | 3 | Pages | 564-567 |
| Mgi Jnum | J:14810 | Mgi Id | MGI:62971 |
| Citation | Kelly CD, et al. (1993) Gene for carbonic anhydrase related protein (CARP): assignment to mouse chromosome 4. Mouse Genome 91(3):564-567 |
| abstractText | Full text of Mouse Genome contribution: GENE FOR CARBONIC ANHYDRASE RELATED PROTEIN (CARP): ASSIGNMENT TO MOUSE CHROMOSOME 4. C.D. Kelly, J. Peters1, N.D. Carter. Medical Genetics Unit, St. GeorgeÕs Hospital Medical School, London SW17 ORE and 1MRC Radiobiology Unit, Chilton, Didcot, Oxon, OX11 ORD, U.K. Introduction The expression of carbonic anhydrase related protein (CARP) cDNA clones has been shown to be located specifically to the Purkinje cells of the mouse cerebellum(1). A number of mouse mutants related to abnormalities or degeneration of Purkinje cells have been described, for example the lurcher, reeler and staggerer phenotype (2). Therefore the localised cerebellar expression of CARP is intriguing and raises the possibility that CARP may be a candidate gene for one of these mutants. As most of the neurological mutants have been mapped it was of interest to locate CARP within the mouse genome. In this study, a mouse CARP cDNA (length 2kb)(l) was hybridized to digested mouse genomic DNA in order to identify a RFLV for C57BL/6J and DBA/2J and then determine the strain distribution pattern in the BXD RI strains, derived from C57BL/6J and DBA/2J. In line with standard nomenclature we now designate the locus Carp. Materials and Methods Genomic DNA was obtained from the Jackson Lab, Bar Harbor Maine or by standard phenol extraction from liver. DNA was digested according to manufacturers' directions. Southern blotting was carried out as described previously(3) and using Zeta-Probe GT blotting membrane (BioRad). Hybridization was carried out overnight at 65 degrees C using 32P-dCTP labelled Carp cDNA. Results and Discussion Three parental/control genomic DNAs from M. spretus, C57BL/6J and DBA/2J were screened for RFLVs using the following restriction enzymes: MspI, BamHI, EcoRI, TaqI, XbaI, RsaI, PvuII, PstI. Digestion of the genomic DNAs with RsaI demonstrated a variation with bands at 2.7kb (C57BL/6J and M. spretus) and 1.7kb (DBA/2J) (Fig 1). Genomic DNA from the set of 26 RI strains of the BXD set was then diegested with RsaI and screened for the 2.7kb/l.7kb variation using the Carp cDNA as a probe (Table 1). Typings for Mos, D4Bir3 and Mtv-14 were taken from refs: 4, 5, 6 and 7 and the results are shown in Table 1. Estimates of recombination frequency and 95% confidence limits were taken from ref 8. No recombinants were found between Carp and D4Bir3 indicating close linkage. The 95% upper confidence limit of the recombination frequency between Carp and D4Bir3 is 4.13%. Linkage was also evident between Carp and the previously described loci Mos(4) and Mtv-l4(7). There are 4 recombinants between Carp and Mos in 26 RI strains giving a recombination frequency of 5.00% with lower and upper 95% confidence limits of 1.17% and 18.28%. There are three recombinants between Carp and Mtv-14 in 26 RI strains giving a recombination frequency of 3.49% with lower and upper 95% confidence limits of 0.64% and 13.76%. These results suggest that Carp lies at the proximal end of Chr 4 between Mos and Mtv-14. Since Carp is located in a conserved segment between mouse and human it is predicted that the human homologue is located on human chromosome 8q(9). An extinct mutant with neurological effects, wd, waddler is also located on proximal Chr 4. Although wd is shown proximal to Mos on consensus maps (9, 10) the position of wd is uncertain as it was mapped in a cross in which the closest marker used was b, 31cM from wd (11). Thus Carp may be a candidate gene for wd. TABLE 1. SDPs of Carp in the BXD RI strains together with SDPs of close markers B-C57BL/6J; D=DBA/2J; X-Crossover BXD strain distribution Locus: Mos; 1: B; 2: B; 5: B; 6: B; 8: B; 9: D; 11: D; 12: B; 13: D; 14: B; 15: D; 16: D; 18: D; 19: D; 20: B; 21: B; 22: B; 23: B; 24: D; 25: D; 27: B; 28: B; 29: B; 30: D; 31: B; 32: D. Locus: D4Bir3; 1: B; 2: B; 5: B; 6: B; 8: B; 9: B; 11: D; 12: B; 13: D; 14: B; 15: D; 16: B; 18: D; 19: D; 20: B; 21: B; 22: D; 23: B; 24: D; 25: D; 27: D; 28: B; 29: B; 30: D; 31: B; 32: D. Locus: Carp; 1: B; 2: B; 5: B; 6: B; 8: B; 9: B; 11: D; 12: B; 13: D; 14: B; 15: D; 16: B; 18: D; 19: D; 20: B; 21: B; 22: D; 23: B; 24: D; 25: D; 27: D; 28: B; 29: B; 30: D; 31: B; 32: D. Locus: Mtv-14; 1: D; 2: B; 5: B; 6: D; 8: B; 9: B; 11: D; 12: B; 13: D; 14: B; 15: B; 16: B; 18: D; 19: D; 20: B; 21: B; 22: D; 23: B; 24: D; 25: D; 27: D; 28: B; 29: B; 30: D; 31: B; 32: D. Acknowledgements. We thank Dr Jean-Louis Guenet for help and advice in the early phase of this work, Dr Kikuya Kato for Carp clone and Dr Ben Taylor for his help in analysing the BXD data. References 1. Kato, K. (1990) F.E.B.S. letters 271, 137-140. 2. Green, M.C. (1989) Genetic variants and strains of the laboratory mouse (eds. M.F. Lyon and A.G. Searle, Oxford). pp. 12-403. 3. Maniatis, T. et al (l989) Molecular cloning - a laboratory manual. Second Edition. 4. Propst, F. et a1 (1989) Genomics 5, 118-123. 5. Birkenmeier, E.H. et a1 (1992) Mammalian Genome 3, 537-545. 6. Birkenmeier, E.H. et a1 (1993) Mammalian Genome 4, 133. 7. Lee, B.K. and Eicher, E.M. (1990) J Virol 64, 4568-4572. 8. Silver, J. (1985). J. Hered. 76, 436-440. 9. Lyon, M.F. & Kirby, M.C. (1993) Mouse Genome 91, 40-80. 10. Hillyard, A.L., et al (1993) Mouse Genome 91, 15-39. 11. Yoon, C.H. (1961) J. Hered. 52, 279-81. Figure 1: (Legend). RFLVs detected with Carp/cDNA Genomic DNA's were digested with Rsal. Key: a = DBA/2J, b = DBA/2J, c = C57BL/6J, d = C57BL/6J, e = BXD 32, f = BXD 31. Variation was scored as presence of a 1.7kb band. |
