POLIMORFISME SINGLE NUCLEOTIDE POLYMORPHISM EXON 9 GEN LEPR PADA AYAM HIBRIDA F2 KAMBRO (Gallus gallus gallus, Linn.1758)

I WAYAN S M, Prof. Dr. Budi Setiadi Daryono, M.Agr.Sc.

2019 | Skripsi | S1 BIOLOGI

Implementasi metode T-ARMS PCR dalam deteksi single nucleotide polymorphisms (SNPs) gen LEPR sampel DNA ayam belum pernah dilakukan. Riset ini bertujuan dalam merancang protokol spesifik deteksi SNPs ekson 9 gen LEPR dan mendeteksi ekspresi gen LEPR atau SNPs LEPR pada sampel DNA ayam Pelung, F1 Pelung, Layer, Broiler Cobb 500, ayam F1 Kambro dan ayam F2 Kambro menggunakan metode T-ARMS PCR. Penentuan derajat korelasi gen LEPR terhadap Bobot Tubuh (BT) dan Produtivitas Telur (PT) pada populasi ayam F1 Kambro dan F2 Kambro. Parameter fenotip kualitatif menunjukkan enam kelompok variasi fenotip tersegregasi dibandingkan ayam F1 Kambro. Pertumbuhan bobot ayam F2 Kambro mencapai 753,36± 155,31 gram dalam 7 minggu tidak signifikan terhadap ayam F1 Kambro disebabkan adanya inbreeding depression (Fx = 25%, LI = 4,925%) dan mutasi transversi alel A LEPR. Protokol spesifik deteksi SNPs ekson 9 gen LEPR dengan metode T-ARMS PCR dapat mendeteksi mutasi C127A LEPR dengan rasio IP : OP 10:1 pmol/µM, konsentrasi template DNA ayam 100 ng/µL dengan temperatur annealing 55,7 C/30s. Mutasi transversi alel A SNPs ekson 9 LEPR terdeteksi pada sampel DNA ayam betina F1 Kambro (80%), ayam jantan F2 Kambro (20%), ayam betina Broiler Cobb 500 (75%). Mutasi tersebut tidak terdeteksi pada sampel DNA ayam Layer, ayam Pelung Blirik Hitam dan ayam F1 Pelung.

The implementation of the T-ARMS PCR method in the detection of single nucleotide polymorphisms (SNPs) in the LEPR gene in chicken DNA samples has never been conducted. This research aims to design a specific protocol for exon 9 LEPR gene SNPs detection and detect LEPR gene expression or LEPR SNPs in Pelung chicken samples, F1 Pelung, Layer, Broiler Cobb 500, F1 Kambro chicken and F2 Kambro chicken using the T-ARMS PCR method. Determination of LEPR gene correlation degree on Body Weight (BT) and Egg Productivity (PT) in F1 Kambro population and F2 Kambro. Qualitative phenotype parameters showed six groups of segregated phenotypes compared to F1 Kambro chicken. Growth of F2 Kambro chicken weight reached 753.36 ± 155.31 grams in 7 weeks was not significant for F1 Kambro chicken due to inbreeding depression (Fx = 25%, LI = 4.925%) and transversion of A LEPR allele mutations. Specific protocol detection of exon 9 LEPR gene SNPs using the T-ARMS PCR method can detect C127A LEPR mutations with IP: OP ratio 10:1 pmol/µM, chicken DNA template concentration of 100 ng/µL with annealing temperature of 55.7 C / 30s. The transversion mutation of C127A of LEPR exon 9 SNP were detected in DNA samples of F1 Kambro hens (80%), F2 Kambro roosters (20%), Broiler Cobb 500 hens (75%). The mutations were not detected in Layer, Pelung Blirik Hitam chicken and F1 Pelung populations.

Kata Kunci : F2 Kambro, F1 Kambro, ARMS-PCR, genotyping, C127A