Evaluation of AMPK Genes as Candidates for Production Traits in Buffalo Breeds of Pakistan

Abstract The 5’-activated protein kinase (AMPK) also called PRKA is a heterotrimeric protein complex that regulates cellular metabolism in response to nutritional stress in mammals. Total length of AMPK gene is 6500 base pairs. AMPK γ3 has 884 base pairs comprises of exon 3, intron 3 and exon 4. AMPK γ3 gene was studied on DNA from three buffalo breeds (Nili, Ravi and Nili Ravi) of Pakistan. Amplification of this region was done from position 918 in exon to position 1802 just downstream of exon 4, using primers U1A-GAGCAAGGAGACAGCACTTCA (918-938) and U2B-ACCTGTAGCATGGTGTC GAAGA (1781-1802) with Amplictaq Gold, at annealing temperature 60oC. Sequencing of PRKA γ3 UP SEQ1 5’-GACCTCAGCATCCAGGCT and PRKA γ3 UP SEQ2 5’-GACAGTAACTCCATCTTCCA was done. The results showed that at position 1069, Allelic frequency of C/T was 0.35, T/T was 0.05 and frequency of C/C was 0.40. At position 1343, frequency of A/G was 0.1 and A/A was 0.9. At 1381 position, frequency of C/T was 0.35, T/T was 0.05 and C/C was 0.6. At position 1488, the A/G was 0.35, A/A was 0.05and G/G was 0.6. The detected SNP at 1343 is A/G which is the ancient haplotypes of PRKAG3 in Nili Ravi buffalo breeds from Pakistan. These SNPs in AMPK gene can be used as marker for efficient utilization of energy in these three buffalo breeds of Pakistan that will increase the economic returns to farmers.