بررسی چندشکلی ژن بتالاکتوگلوبولین گوسفند به روش PCR-RFLP

In modern programmes of animal breeding, the polymorphisms of the milk proteins can be used as marker systems. Beta-Lactoglobulin is the major milk whey protein in the ruminants. The BLG coding gene located on ovine chromosome 3. This protein, synthesis in the mammary glands during pregnancy and the lactation stages. Studies have indicated that this protein is polymorphic in the many breeds of sheep. This is the result of a single base pair substitution in the Beta-Lactoglobulin gene that also rises to the RsaI restriction fragment length polymorphism (RFLP). The aim of this work was to analyze the genotype distribution of Beta-Lactoglobulin in sheep. Blood samples were supplied from 142 sheep of the 5 breeds (Ghezel, Afshari, Moghani, Makoii and Arkharmerino). Genomic DNA was extracted from the 200ul blood sample according to Boom et al. (1989) method modified by Shaikhayev (1995). The Gel monitoring and the spectrophotometeric methods were used for determination of the DNA quality and quantity. the Primers BLG5 and BLG3 amplified a 452 bp fragment from the exon II of the ovine Beta-Lactoglobulin gene. the Products of the amplification were recognized by the electrophoresis on the 1% agarose gel stained with ethidium bromide. The RsaI enzyme was used for restriction of the PCR products. The digested products were separated by the electrophoresis on the 8% nondenaturant polyacrylamide gel and visualized after staining with the ethidium bromide on UV transillumination. The popGen32 software (ver.1.31) was used to estimate the allele and the genotype frequencies, the Hardy-Weinberg equilibrium and dendrogram of the genetic distance. The frequency of A-allele in Ghezel, Afshari, Moghani, Makoii and Arkharmerino breeds was 56%, 34%, 36%, 53% and 48% respectively. The populations were in Hardy-Weinberg equilibrium except to Afshari breed. The lowest genetic distance was observed between Moghani and Afshari breeds and the highest genetic distance between Ghezel and Afshari breeds. The results of this study indicated that PCR-RFLP is an appropriate tool for evaluating genetic variability in sheep.