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The single-strand conformation polymorphism(SSCP) is a point mutation screening method based on changes in the secondary structure in a defined single-stranded DNA fragment caused by a change in sequence. This study was performed to develop the identification technique of Hanwoo meat using PCR-SSCP marker of MC1R gene associated with the coat colors of cattle. Hanwoo, Holstein and imported cattle breeds(Angus, Hereford and Charolais) were used for the detection of PCR-SSCP genotypes of MC1R gene. The specific primers were used to amplify a 138bp and a 390bp of the bovine E-locus(including point mutation) corresponding to positions 318-455 and 318-707, respectively, in the MC1R gene. For SSCP genotype analysis, the PCR products were denatured at 95℃ for 5min, loaded on a 12-15% nondenaturing polyacrylamide gel and detected by silver staining. When the PCR product of 138bp was separated by gel electrophoresis, two genotypes of E^+/e and e/e were detected only in Hanwoo meat, but E^D/E^D and E^D/e and E^D/E^D genotypes were observed in Holstein meat and Angus breed, respectively. Also, in the SSCP analysis using PCR product of 390bp, Hanwoo meat produced banding pattern of two fragments, while single fragment was observed in Holstein meat and Angus breed. Therefore, breed-specific SSCP markers showing distinct differences between Hanwoo and Holstein and Angus were found by PCR-SSCP analysis. This study shows that the PCR-SSCP technique is relatively simple, fast and a practical tool for determination of MC1R genotypes of cattle breeds when compared with PCR-RFLP method. Therefore, the PCR-SSCP markers of MC1R gene could be used as DNA markers for identification of Hanwoo meat from Holstein and Angus meats.
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