Identification and characterization of a microsatellite marker within murine Brca2 gene

Identification and characterization of a microsatellite marker within murine Brca2 gene Mammalian Genome 8, Brief Data Reports 79 5. Kitamura, N., Kitagawa, H., Fukushima, D., Takagaki, Y., Miyata, T., Nakanishi, S., (1995). J. Biol. Chem. 260, 8610--8617. 6. Kitamura, N., Takagaki, Y., Furuto, S., Tanaka, T., Nawa, H., Nakani- shi, S. (1983). Nature 305, 545-549. 7. Harlizius, B., Hetzel, J., Barendse, W. (1995). Mamm. Genome 6, 486--483. 8. Moody, D.E., Pomp, D., Barendse, W. (1995). Anim. Genet. 26, 45- 9. Solinas-Toldo, S., Lengauer, C., Fries, R. (1995). Genomics 27, 489- 10. Threadgill, D.S., Womack. J.E. (1991). Genomics 11, 1143-1148. 11. Irie, Y., Tatsumi, K., Notomi, T., Endo, Y., Onogi, S., Miyai, K., Amino, N. (1993). Human Genome Mapping Workshop 93, 57. 12. Qumsieh, M.B., Valentine, M.B., Suttle, D.P. (1992). Genomics 5, Fig. 2. Polymorphism detection with PCR. Shown here are the PCR prod- 160-162. ucts amplified from the genomic DNA of Mus musculus, M. spretus, and 13. van Rens, G.L.M., Raats, J.M.H., Driessen, H.P.C., Oldenburg, M., M. musculus/M, spretus hybrid mice and separated on a 4% agarose gel in Wijnen, J.T., Meera Khan, P., de Jong, W.W., Bloemendal, H. (1989). lx TBE buffer. Gene 78, 225-233. 14. Cox, D.R., Kawashima, H., Vora, S., Epstein, C.J. (1984). Cytogenet. was found to be polymorphic between Mus musculus and Mus Cell Genet. 37, 441--442. spretus species (Fig. 2). Such a polymorphic marker within in a 15. Hawkins, J.W., Van Keuren, M.L., Piatigorshy, J., Law, M.L., Patter- putative tumor suppressor gene can be very useful in detecting loss son, D., Kao, F.T. (1987). Hum. Genet. 76, 375-380. of heterozygosity in tumor samples. Acknowledgments: The authors thank Lois J. Maltais of The Jackson Laboratory for help in choosing the locus symbol that meets the nomecla- Identification and characterization of a ture guidelines. This research was supported by grants from National In- stitute of Health and Howard Hughes Medical Institute. A. Bradley is an microsatellite marker within murine Associate Investigator with HHMI. Brca2 gene References 1. Wooster, R., Bignell, G., Lancaster, J., Swift, S., Seal, S., Manglon, J., S.I~ Sharan, A. Bradley Collins, N., Gregory, S., Gumps, C., Micklems, G., Barfoot, R., Ham- oudi, R., Patel, S., Rice, C., Biggs, P., Hashim, Y., Smith, A., Connor, Howard Hughes Medical Institute, Department of Molecular and Human F., Arason, A., Gudmundsson, J., Ficenec, Kelsell, D., Ford, D., Tonin, Genetics, Baylor College of Medicine, One Baylor Plaza, P., Bishop, D.T., Spurt, N.K., Ponder, B.A., Eeles, R., Peto, J., Devilee, Houston, Texas 77030, USA P., Cornelisse, C., Lynch, H., Narod, S., Lenoir, G., Egilsson, V., Bark- adottir, R.B., Easton, D.F., Bentley, D.R., Futreal, P.A., Ashworth, A. Received: 28 August 1996 / Accepted: 5 September 1996 and Stratton, M.R. (1995). Nature 378, 789-792. 2. Tavtigan, S.V., Simard, J., Rommens, J., Couch, F., S.hattuck-Eidens, Species: Mus musculus D., Neuhausen, S., Merajver, S., Thorlacius, S., Offit, K., Stoppa- Locus symbol: D5Bayl Lyonnet, D., Belanger, C., Bell, R., Berry, S., Bogden, R., Chen, Q., Database deposit information: Genebank accession Number: Davis, T., Dumont, M., Frye, C., Hattier, T., Jammulapati, S., Janecki, U66168 T., Jiang, P., Kehrer, R., Leblanc, J.-F., Mitchell, J.T., McArthur- Molecular reagent: Forward primer, 5' CIT GTC TGG CAA Morrison, J., Nguyen, K., Peng, Y., Samson, C., Schroeder, M., Snyder, CAA ACG CA3'; reverse primer, 5' CAA GAG AAG GTG GTT S.C., Steele, L., Stringfellow, M., Stroup, C., Swedlund, B., Swensen, CAG AGY. B., Teng, D., Thomas, A., Tran, T., Tranchant, M., Weaver-Feldhaus, J., Wong, A.K.C., Shizuya, H., Eyfjord, J.E., Cannon-Albright, L., Labrie, Allele detection condition: PCR reaction: 50 ng of total genomic F., Skolnick, M.H., Weber, B., Kamb, A., and Goldgar, D.E. (1996). DNA, lxPCR buffer (Perkin Elmer), 1.0 mM MgCI2, 200 p.M Nature Genet. 12, 333-337. dNTPs, 1 I~M of each primer, and 2.5 units of Taq DNA polymer- ase in 50 i~1 total reaction volume. PCR conditions: 30 cycles of Using SSCP to facilitate mapping 94~ 1 rain; 55~ 1 min; 72~ 2 rain. Discussion: The microsatellite marker within the mouse homolog microsatellite loci of human breast and cancer susceptibility gene BRCA2 was iden- tified in an intron corresponding to human BRCA2 intron number J.L Williams, 1 I. Oisaker, 2 V.M. Teres 1 14 [1,2]. The microsatellite marker contains about 20 CA repeats (Fig. 1). Under the PCR conditions described above, the primers 1Division of Molecular Biology, Roslin Institute (Edinburgh), were used to amplifty 225 bp of mouse genomic DNA. The marker Midlothian, Scotland EH25 9PS, UK 2Department of Morphology, Genetics and Aquatic Biology, Division of Genetics, Norwegian College of Veterinary Medicine, P.O. Box 8146 Correspondence to: A. Bradley Dep. N-0033, Oslo, Norway Received: 15 July 1996 / Accepted: 21 August 1996 1 CCGGAGAGAG AGTACTTCAG TATGCTTGTC TGGCAACAAA CGCACGTACT Species: Bos tarus 51 GTGTGTGTGC GTGCATGTGT GTGTGTGTGT GTGTGTGTGT GTGTGTGTGT Locus names: RI5 (D7S33), RI13 (D22S27), RI15 (D28S24), RI918 (D2S32) 101 GTGTGCGTGT GCGTGTGCCT GTGTGTTGAG AGACAGAGAT CAGGTGACTG Map position: See Table 1. 151 ATTTGCTCTT GTTAGCATTT ACACCACAGA ACGATACCAC AACATCCAAG Method of mapping: Segregation of alleles for the four markers was confirmed by typing the 347 individuals comprising 15 sire 201 GCTGTCGTTA TTCCTTACAA GTGTGGCCCC TCCCGCTCTG AACCACCTTC families of the International Bovine Reference Panel (IBRP) [1]. 251 TCTTGGCGAC CTGGAGGGGC TTACATGCTA GTAGCTCAGT CTTCTCTTCC Markers were assigned to bovine chromosomes by two-point link- 301 ATTCCAATCC CACCCGCCTT age analysis (CRIMAP). Molecular reagents: Oligonucleotide primer sequences are shown Fig. 1. Nucleotide sequence of D5Bayl. The underlined bases represent the polymorphic repeat present within the locus. in Table 1. For RI5, RI13, and RI15 sequences were obtained by Mammalian Genome Springer Journals

Identification and characterization of a microsatellite marker within murine Brca2 gene

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