Abstract: The aims of this research was to identifying myostatin gene (MSTN) polymorphism and the it is association with body weight, sex and birth weight in Padjadjaran sheep, blood samples were collected from the jugular vein 3 ml of each head, used kit DNA genomic samples of whole blood 2 ml DNA isolation, to identify candidate gene used the Ensenbl database, three pairs of primers were designed to amplify exons 1, 2 and 3 of MSTN gene. The 25 μl volume contained 50 ng of genomic DNA, 12.5 μl 2x Reaction mix, 0.5 μM of each primer, and 0.5 units of Taq DNA polymerase. The cycling protocol was 5 min at 95°C, 30 cycles of denaturation at 94°C for 30 second, annealing at 58-68°C for 1 second and extended at 72°C for 1 second, with final extension at 72°C for 3 second. All samples sent to Malaysia for sequencing and used BioEdit program to identify nucleotide substitution and sequenced results compared with MSTN gene reference coding regions. Calculation of genotypes, allele’s frequencies, mean expected and observed heterozygosities, and Chi-square test was performed. Body weight and body measurements were used for statistical analysis by GLM procedure and the LSD (SAS version 9.0). Results showed that exon 1 of c.101G>A position was dimorphic and c.120insA genotype was monomorphic, and c.960delG locus was polymorphic with genotype frequencies of GG 96.872% and GC 5.128% respectively this locus did not show Hardy-Weinberg equilibrium (P <0.05). SNP is an AAG →TCG and AAG → GCG substitutions allocated in the 3rd exon, these variations only detected in two individuals were found to be heterozygous for the c.960delG. An association was revealed between c.960delG,gender and birth weight were (p = 0.205) and (p = 0.042) respectively, and body weight (P>0.001). It can be concluded that, although MSTN polymorphism effect on body weight, birth weight and sex, further study needs to be conducted and involving many more sheep of Padjadjaran breed including the other Indonesian sheep to confirm these observations.

 

Key words: identification, MSTN gene, Padjdjaran sheep and RT-PCR