BRIEF REPORTS 909 extending known homology between mouse chromosome 1 and human chromosome 2. The five members of subfamily 2 are probably derived from independent genes (4). Southern blot analysis of mouse geno- mic DNA, using androsterone UGT, indicated the presence of several independent but homologous UGT2 genes (4). Due to the difficulty in isolating suitable specific cDNA probes from the UGT2 members, we have employed the polymerase chain reaction (PCR) in the chromosomal mapping of the steroid UGT gene encoding a bile acid UGT cDNA (Hlug25; (5)). This technique has been successfully used to map the chromosomal location of members of other gene families (1). Two oligonucleotide primers were selected from regions of the published Hlug25 cDNA sequence (see (5)) that showed the least sequence homology to the other UGT2 cDNAs iso- lated (3). The amplification cycle was 94°C for 1 min, 55°C for 1 min, and 72°C for 1 min 30 s and 25 cycles were performed. Analysis of the PCRproducts was carried out by gel electropho- resis. The Hlug25 primers were shown to specifically amplify human genomic DNA and Hlug25 cDNA but not rodent geno- mic DNA or other human UGT2 cDNAs isolated (Fig. 1). In addition, restriction digestion of the amplified products con- firmed that an exonic region of the Hlug25 gene had been spe- cifically amplified. Analysis of a panel of 23 human/rodent somatic cell hybrid lines by PCR using the Hlug25-specific primers indicated the presence of the Hlug25 gene in 11 of these lines. A concordance/discordance analysis revealed a 100% concordance and a 0% discordance of the Hlug25 gene for human chromosome 4. A UGT gene encoding a transferase catalyzing steroid glu- curonidation has been localized to mouse chromosome 5 (see (3)). This is of interest since there is a considerable region of homology between mouse chromosome 5 and human chromo- some 4 (7). The mouse gene mapped is not the exact equivalent of the human gene described in this report, the enzymes hav- ing different substrate specilities; however, it will be interest- ing to see if the various UGT2 genes are clustered on mouse chromosome 5 and human chromosome 4. PCR mapping using specific sequences from individual gene products will facilitate chromosomal localization of other sub- family 2 members to determine whether independent genes exist on the same or different chromosomes, thereby providing further knowledge of the evolution of this gene family. ACKNOWLEDGMENTS We thank the Medical Research Council and the Wellcome Trust for supporting this research. G. Monaghan holds an HGMP student- ship. We are grateful to Sue Povey and Nigel Spurr for the hybrid lines and to Steve Jeremiah and Andrea Bryan for technical assistance. REFERENCES 1. Abbott, C., West, L., Povey, S., Jeremiah, S., Murad, Z., Discipio, R., and Fey, G. (1989). 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S., Moss, J. E., Taylor, B. A., Burchell, B., and Wolf, C. R. (1991). Mapping genes encoding drug-metabolizing en- zymes in recombinant inbred mice. Genomics 11: 309-316. Nadeau, d. H., and Reiner, A. H. (1989). Linkage and synteny homologies in mouse and man. In “Genetic Variants and Strains of the Laboratory Mouse” (M. F. Lyon and A. G. Searle, Eds.), 2nd ed., pp. 506-536, Oxford Univ. Press, London/New York. Ritter, J. K., Crawford, J. M., and Owens, I. S. (1991). Cloning of two human liver bilirubin UDP-glucuronosyltransferase cDNAs with expression in COS-1 cells. J. Biol. Chem. 266: 1043-1047. Wooster, R., Sutherland, L., Ebner, T., Clarke, D., Da Cruz, E., Silva, O., and Burchell, B. (1991). Cloning and stable expression of a new member of the human liver phenol/bilirubin UDP-glu- curonosyltransferase cDNA family. Biochem. J. 278: 465-469. Detection of Mutations in the Factor VIII Gene Using Single-Stranded Conformational Polymorphism (SSCP) Effrosini P. Economou, Haig H. Kazazian, Jr., and Stylianos E. Antonarakis’ Center for Medical Genetics, Departments of Pediatrics and Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205 Received November 21, 1991; revised January 31, 1992 Hemophilia A is an X-linked disorder of blood coagulation due to deficiency of factor VIII. The gene for factor VIII con- tains 26 exons and spans 186 kb (5). Many mutations in the factor VIII gene have been characterized in patients with He- mophilia A. (1). We used single-stranded conformational poly- morphism (7) (SSCP) to screen for mutations in a given PCR- amplified DNA product. Four exons and their splice consensus sequences were studied in DNA from 168 patients from the Hemophilia Centers of the Orthopedic Hospital of Los An- geles (Dr. C. Kasper), Vanderbilt University (Drs. R. Janco and J. A. Phillips), Tokyo Medical College (Dr. M. Fujimaki), University of Bonn (Dr. K. Olek), St. Sophia Hospital, Ath- ens, (Dr. S. Aronis), and the DNA Diagnosis Laboratory of The Johns Hopkins Hospital. Exons II, 16, 17, and 24 and their consensus splice se- quences were amplified after PCR [see (7) for primers and conditions of the PCR reaction]. One of the two primers was end-labeled with 100 &i [-y-32P]ATP at 6000 Ci/mmol with ’ To whom reprint requests should be addressed. CENOMICS 13,90%911 (19%) 088%7543/92 $5.00 Copyright 0 1992 by Academic Press, Inc. All rights of reproduction in any form reserved.