Detection of polymorphism in upstream regulatory region of calpain and calpastatin genes and its association with body weight and fat-tail characteristics traits in Lori-Bakhtiari sheep

The aim of the present study was to investigate allelic diversity in upstream regulatory region of calpastatin and calpain genes, bioinformatics analysis of studied genes and association of allelic variants with body weight and fat-tail traits in Lori-Bakhtiari sheep.
One hundred and fifty blood samples were collected randomly and DNA was extracted by modified salting out method. The specific primer pairs were designed using Oligo 7 software for amplification of fragments with lengths of 245 and 225 bp from upstream regulatory region of calpastatin and calpain genes, respectively. After PCR, the amplified products of calpastatin and calpain marker sites were subjected to endonuclease digestion with PstI and HaeIII enzymes, respectively. Due to the monomorphic of RFLP results, the SSCP technique was used for more accurate assessment of amplified fragments. After genotyping, one sample from each banding patterns of calpain gene was sequenced. The obtained sequences were investigated for identification of motifs involved in gene regulation and also mutations in different banding patterns by DNASIS MAX and BioEdit softwares. Association between observed banding patterns and body weight traits at birth, 3, 6 and 12 months of age and weight, upper bound, lower bound and the height of fat-tail was investigated by Glimmix procedure of SAS (version 9.1) software.
Three banding patterns of A, B, and C with frequencies of 66, 9, and 25% were observed in Lori-Bakhtiari samples, respectively. The upstream region of calpastatine gene was monomorph in present study. Statistical analysis showed significant association between banding patterns of calpain marker site and body weight at birth in Lori-Bakhtiari sheep (P According to the polymorphic pattern in upstream regulatory region of calpain locus and its significant association with birth weight, this marker site may be used in breeding programs to improve meat quality in sheep.