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Characterization of transcribed ovine lymphocyte antigen (OLA) class I genes by single strand conformational polymorphism (SSCP) and sequence analysis

  • D. Miltiadou (a1), K. T. Ballingall (a2), S. A. Ellis (a3) and D. J. McKeever (a1) (a2)

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The Major Histocompatibility Complex (MHC) class I genes encode polymorphic cell surface glycoproteins that initiate cytotoxic immune responses by presenting processed antigenic peptides to CD8+ T cells. Due to their functional role and polymorphic nature, MHC class I genes have been used in disease association and vaccine development studies in many species. A prerequisite for such studies is the availability of animals of defined MHC haplotypes. However, the ovine MHC (OLA) class I genes have been studied largely by serology and very limited information is available at the DNA level (Dietert R., 1996). In this study, we aimed to identify the number of transcribed OLA class I genes in two Blackface rams and to obtain sequence data for exons 2 and 3. These data will be used to establish an allele specific PCR based typing system to genotype a cohort of progeny for each ram. This will enable allocation of alleles to individual haplotypes and inform a breeding strategy for the generation of MHC defined animals for functional and disease association studies.

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Ainsworth, P. J., Surh, L. C. and Coulter-Mackie, M. B. 1991. Diagnostic single strand conformational polymorphism, (SSCP): a simplified non-radioisotopic method as applied to a Tay-Sachs B1 variant. Nucleic Acids Research 19 (2):405406.
Dietert, R. R. 1996. The ovine major histocompatibility complex . In The Major Histocompatibility Complex Region of Domestic Animals. Edited by Schook, LB. and Lamont, SJ. CRC Press. Chapter 4: 6598.
Hawker, J. K. and Billadello, J. J. 1993. A multiplex polymerase chain reaction colony miniprep. Biotechniques 14 (5):764

Characterization of transcribed ovine lymphocyte antigen (OLA) class I genes by single strand conformational polymorphism (SSCP) and sequence analysis

  • D. Miltiadou (a1), K. T. Ballingall (a2), S. A. Ellis (a3) and D. J. McKeever (a1) (a2)

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