Analysis of the sheep MHC using HLA class I,II, and C4 cDNA probes |
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Authors: | Dr. Patrick Chardon Marek Kirszenbaum Philippa R. Cullen Claudine Geffrotin Charles Auffray Jack L. Strominger Daniel Cohen Marcel Vaiman |
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Affiliation: | (1) Laboratoire de Radiobiologie Appliquée, Institut National de la Recherche Agronomique, C.N.R.Z., 78350 Jouy-en-Josas, France;(2) Laboratoire de Radiobiologie Appliquée, Commissariat à l'Energie Atomique, I.P.S.N.-S.P.E., 78350 Jouy-en-Josas, France;(3) Laboratoire de Génétique Biochimique, INRA, 78350 Jouy-en-Josas, France;(4) Institut d'Embryologie, CNRS, 94130 Nogent s/Marne, France;(5) Department of Biochemistry and Molecular Biology, Harvard University, 02138 Cambridge, Massachusetts;(6) Hôpital Saint Louis, INSERM U93, 75010 Paris, France |
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Abstract: | Four cDNA probes for the human major histocompatibility complex (MHC) were used to investigate the sheep MHC, in conjunction with serological typing for ovine lymphocyte antigen (OLA). Lymphocytes from a family (two parents and five offspring) of Romanov sheep were subjected to genomic DNA digestion by the restriction endonuclease Eco RI, followed by gel electrophoresis. A single Southern blot representing all seven individuals was then consecutively hybridized with the class I, alpha-DC, beta-DR, and C4 probes, which were originally designed to identify HLA class I, class II (DC and DR), and C4 products, respectively. Using each of the three class I/class II probes, several bands showing DNA polymorphism were detected. The segregation of these bands in the five offspring exactly paralleled the OLA haplotype segregation established by serological typing. A further eight individuals carrying haplotypes which were phenotypically identical to those in the above-mentioned family showed bands in the corresponding positions when tested with the same three probes. Using the C4 probe, no polymorphism was detected in these fifteen individuals.Abbreviations used in this paper MHC major histocompatibility complex - OLA ovine lymphocyte antigen - kbp kilobase pair(s) - MLR mixed lymphocyte reaction - RFLP restriction fragment length polymorphism |
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