Copyright 0 1990 by the Genetics Societyof America A Genetic Linkage Map of Mouse Chromosome 10: Localization of Eighteen Molecular Markers Using a Single Interspecific Backcross Monica J. Justice,* Linda D. Siracusa,* DebraJ. Gilbert,* Nora Heisterkamp,?John Groffen,+ Kiran Chada,*Colleen M. Silan,* Neal G. Copeland* and Nancy A. Jenkins* Mammalian Genetics Laboratory, ABL-Basic Research Program, NCI-Frederick Cancer Research Facility, Frederick, Maryland 21 701,'Section of Molecular Diagnosis, Department of Pathology, Childrens Hospital of Los Angeles, Los Angeles, Calqornia 90027, and "Departmentof Biochemistry, University of Medicine and Dentistry of New Jersey, Robert WoodJohnson Medical School, Piscataway, New Jersey 08854 Manuscript received January2, 1990 Accepted for publication April20, 1990 ABSTRACT Interspecific mouse backcross analysis was used to generate a molecular genetic linkage map of mouse chromosome 10. The map locations of the Act-2, Ahi-I, Bcr, BraJ Cdc-2a, Col6a-1, Co16a-2, Cos-I, Esr, Fyn, Gli, Ijg,Igfl, Myb, Pah,pgh", Ros-1and SlOOb loci were determined. Theseloci extend over 80% of the genetic length of the chromosome, providing molecular access to many regions of chromosome 10 for the first time. The locations of the genes mapped in this study extend the known regions of synteny between mouse chromosome 10 and human chromosomes 6, 10, 12 and 21, and reveal a novelhomology segment between mouse chromosome IO and human chromosome 22. Several loci may lie close to, or correspond to, known mutations. Preferential transmission of Mus spretus-derived alleles was observed for loci mapping to the central region of mouse chromosome 10. HE development of mouse molecular genetic AVNERet al. (1988)l. In a laboratory setting, the wild T linkage maps has proved invaluable forthe mouse species Mus spretus will interbreed with inbred structuraland functional characterization of the laboratory mouse strains to produce fertileF1 females mouse genome. First, molecular genetic linkage maps and sterile F1 males (BONHOMMEet al. 1984). The F1 have identified molecular markers that represent pre- females can then be backcrossed to generate the NB viously isolated mouse mutations (CHABOTet al. 1988; progeny used in mapping studies. GEISSLER,RYAN and HOUSMAN1988; BALLING, An IB involving the inbred laboratorymouse strain DEUTCHand GRUSS1988). Second, molecular genetic C57BL/6J and M. spretus was used to create a multil- linkage maps have been utilized to determine whether ocus molecular genetic linkage map of mouse chro- newly identified genes or viral integration sites are mosome 10. Mutations previously mapped to mouse homologous to known genes or mutations (BUCHBERG chromosome 10 include many loci affecting coat et al. 1988; MUCENSKIet al. 1988; SOLAet al. 1988; color, hematopoiesis, neural development, behavior, BARTHOLOMEWet al. 1988). Third, molecular genetic bonedevelopment and growth(summarized by linkage maps have been used for comparative map- GREEN1989). We were interested in creating a mo- ping between mouse and human (or other) genomes lecular geneticmap of chromosome IO to localize [reviewed by NADEAU(1989) and SEARLEet al. (1989)]. proto-oncogenes, common sites of viral integration, Comparative mapping can ultimately lead to the iden- growthfactors, growth factor receptors andother tification of mouse models of human diseases (BRUE- genes affecting cell differentiation and growth. Prior TON et al. 1988; WINTER1988; RYDER-COOKet al. to this study, only three molecular markers had been 1988; GLASERand HOUSMAN1989). positioned on chromosome IO. The Myb proto-onco- One of the most powerful methods for mapping a gene had been mapped to chromosome 10 by in situ large number of molecular markers in the mouse is hybridization and mapped relative to Steel (Sl) in interspecific backcrosses (IBs) between distantly re- intraspecific crosses (SAKAGUCHIet al. 1984; TAYLOR lated species of mice [reviewed by GUENET(1986) and and ROWE1989). Phenylalanine hydroxylase (Pah) had AVNERet al. (1988)l. The evolutionarydivergence been mapped to chromosome 10 by IBs and by in situ between species has resulted in an accumulation of hybridization analysis (BODEet al. 1988; LEDLEYet ai. DNA sequence differences (BONHOMMEet ai. 1984) 1988). The autosomal Zincfinger protein(Zfa) had been that facilitates the detection of restriction fragment mapped relative to Myb and Pah in 1Bs (MITCHELLet length polymorphisms (RFLPs) required for mapping al. 1989). Several other loci had been placed on molecular markers (FERRIS, SAGEand WILSON1982; chromosome 10 by somatic cell hybrid analysis, in- ROBERTet al. 1985) [reviewed by GUENET(1986) and cluding Abelson helper virus integration site-I (Ahi-l), Genetics 125 855-866 (August, 1990) 856 M. J. Justice et al. collagen (~l(V1)(Col6a-1), collagen a2(VI) (Co/6a-2), pBluescript SK+ (Cl-BS; CISEKand CORDEN1989) thatwas interferon-y (Ifg), and SlOO protein, ,6 subunit (S100b) a gift from J. L. CORDENUohns Hopkins School of Medicine (Baltimore, Maryland)]. (POIRIER,KOZAK and JOLICOUER 1988; WEIL et al. The probes forcollagen al(VZ) (Col6a-I) (p18;WEIL et al. 1988; NAYLOR,GRAY andLALLEY 1984). In addition, 1988) and collagen aZ(VZ) (CoL6a-2) (pl; CHU et al. 1987) an anonymous mouse probe identifying the Cos-1 lo- were human cDNA EcoRI fragments cloned in pUCl9; both cus had been placed on chromosome 10 by insitu probes were gifts from M.-L. CHU [ThomasJefferson Uni- hybridization (G. RADICE,J. LEEand F. COSTANTINI, versity (Philadelphia, Pennsylvania)]. The anonymous mouse probe (Cos-1) was a gel-purified personal communication). Allof these molecular mouse brain cDNA 1.2-kb EcoRI fragment (p2351 ; G. RAD- markers except Zfa were mapped in our IB relative to ICE, J. LEEand F. COSTANTINI,unpublished results) that was each other and formed the coreof reference loci used a gift from F. COSTANTINI[Columbia University, College of to localize previously unmapped genes. After the ref- Physicians and Surgeons(New York, New York)]. erence loci were placed on mouse chromosome 10 The probe for the estrogenreceptor (Esr) was a human cDNA EcoRI fragment cloned in pBR322 (pOR3; GREENet using a single [(C57BL/6J X M. spretus)F1 X C57BL/ al.1986) that was purchasedfrom the American Type 6J] IB, an actin-related locus (Act-2), the breakfioint Culture Collection (Rockville, Maryland). cluster region gene (Bcr), the Braf transforming gene The probe for the Fyn proto-oncogene (Fyn) was a human (Braf), a cell division cycle control protein gene (Cdc- cDNA Sac1 fragment cloned in pUC18 (KAWAKAMI,PEN- 2a), the estrogen receptor (Esr), the Fyn proto-oncogene NINGTON and ROBBINS1986) thatwas a gift from K. ROBBINS [National Institutes of Health (Bethesda, Maryland)]. (Fyn), the glioblastoma proto-oncogene (Gli), the insulin- The probe for theglioblastoma proto-oncogene (Gli) was a like growth factor-1 gene (Zgfl), a transgenic integra- mousegenomic EcoRI/HindIII fragmentcloned into tion at the pygmy locus (pg"'), and the Ros-l froto- pBluescript (pMGLI-RS; K. W. KINZLER,J. M. RUPPERT oncogene (Ros-1) were mapped to chromosome 10. and B. VOGELSTEIN,unpublished results; KINZLER et al. The results of the IB analysis provide an unambig- 1988)that was agift from B. VOGELSTEIN[The Johns Hopkins Oncology Center (Baltimore, Maryland)]. uous orientation of 18 molecular markers that span The probefor interjeron-y (rfg) was amouse cDNA most of mouse chromosome 10. The map locations of BamHI fragment cloned in pCD (K.-I. ARAI,unpublished the loci compared with the composite genetic linkage results) that was a gift fromH. YOUNG [NCI-Frederick map suggests that several probes may lie close to, or Cancer Research Facility (Frederick, Maryland)] with per- correspond to, known mouse mutations. The results mission from K.-I. ARAI[DNAX (Palo Alto, California)]. The probe for theinsulin-like growth factor-1 gene (Zgfl) also reveal extensive regions of synteny with human was amouse cDNA EcoRI fragment cloned in pBR327 chromosomes 6, 10, 12 and 21, and reveal a novel (pmigfl-2; BELLet al. 1985) that was a gift from G. BELL homology segment with human chromosome 22. [Howard Hughes Medical Institute (Chicago, Illinois)]. The probe for the Myb proto-oncogene (Myb) was a mouse MATERIALSAND METHODS genomic XbaI fragment cloned in pUCl2 (SHEN-ONGet al. 1984) that was a gift from G. SHEN-ONG[National Cancer Mice: The interspecific backcross [(C57BL/6J X M. spre- Institute (Bethesda, Maryland)]. tus)F1 X C57BL/6J] was performed at the NCI-Frederick The probe forphenylalanine hydroxylase (Pah) was synthe- Cancer Research Facility as described by BUCHBERGet al. sized by the polymerase chain reaction amplification (SAIKI (1 988, 1989). et al. 1988) of mouse liver cDNA using synthetic oligode- Probes: The probe for theAbelson helper virus integration oxynucleotides corresponding to nucleotides 474-493 and site-1 (Ahi-1) was a mouse PstI-Hind111 genomic fragment the complement of nucleotides 1254-1273 of the rat phen- cloned in pUCl8 (p2-1; POIRIER, KOZAK and JOLICOUER ylalanine hydroxylase cDNA sequence (DAHL andMERCER 1988) that was a gift from P. JOLICOUER [Clinical Research 1986). Institute of Montreal (Montreal, Quebec, Canada)]. The probe for the transgenic integration at the pygmy The probe for theactin-related locus (Act-2) was a chicken locus (pgha)was amouse genomic fragment cloned in @actin full-length cDNA cloned in pBR322 (82000; CLEVE- pBluescript (XIANG,BENSON and CHADA 1990). LAND et al. 1980) that was a gift from S. H. HUGHES[NCI- Theprobe for