ANALISIS MOLEKULAR GENETIK DOMBA BATUR DENGAN RANDOM AMPLYFIED POLYMORPHISM DNA DAN RESTRICTION FRAGMENT LENGTH POLYMORPHISM GENSITOKROMB
PRAYITNO, Prof. Dr. drh. Wayan Tunas Artama
2013 | Disertasi | S3 BioteknologiDomba atur memiliki ciri morfologi mirip Merino serta perpaduan domba lokal dan Merino. Status genetik domba Batur belum diketahui, karena tidak ada catatan silsilah yang jelas. Identifikasi domba Batur dengan ciri morfologi hasilnya tidak akurat. Melalui analisis dengan Random Amplyfied Polymorphism DNA (RAPD) genom dan Restriction Fragment Length Polymorphism (RFLP) gen sitokromb dapat diketahui kesamaan dan jarak genetik dalam satu dan lintas populasi, status genetik dan silsilah tetua domba Batur. Tujuan penelitian untuk mendapatkan marka RAPD dan RFLP gensitokromb spesifik domba Batur. Analisis RAPD digunakan DNA dari 89 sampel domba Batur, Merino, Garut, Ekor Tipis dan Ekor Gemuk, sedangkan untuk analisis RFLP gensitokromb masing-masing 9 ekor. Analisis RAPD digunakan empat macam primer 10 mer dan satu primer 11 mer serta PCR gensitokromb dengan dua macam primer 25mer. Analisis RFLP gensitokromb digunakan enzim restriksi Hae III. Diidentifikasi pita RAPD dan RFLP gensitokromb mono morfik dan polimorfik. Amplifikasi 89 sampel domba dengan lima macam primer RAPD dihasilkan 3.921 fragmen DNA berukuran 100 sampai 1500 bp. Jumlah pita mono morfik dan polimorfik masing-masing 1.946 dan 2.243. Analisis RFLP gen sitokromb dihasilkan delapan macam fragmen berukuran 118 sampa i232 bp. Jumlah total pita RAPD paling banyak adalah sampel domba Batur (1.382) dan paling sedikit domba Ekor Gemuk (493). Pita mono morfik paling banyak pada sampel domba Batur (729) dan paling sedikit domba Garut (204), sedangkan untuk pita polimorfik paling banyak pada sampel domba Batur (653) dan paling sedikit domba Merino (90). Pita monomorfik serta polimorfik yang spesifik untuk rumpun domba Batur adalah 363 dan 1500 bp, Merino 225 dan 345 bp,Garut 790 dan 530 bp, Ekor Tipis 615 dan 620 bp serta Ekor Gemuk 615 dan 720 bp. Antar individu dalam populasi domba Batur dan Garut, Ekor Tipis dan Ekor Gemuk memiliki kesamaan genetik lebih rendah dibandingkani ndividu dalam populasi Merino. Kesamaan genetik antar individu satu populasi paling tinggi adalah dalam populasi domba Merino, dan paling rendah domba Garut, sedangkan untuk lintas populasi paling tinggi domba Batur dan Merino dan paling rendah Merino dan Ekor Tipis. Jarak genetik paling dekat adalah populasi domba Batur dan Merino dan paling jauh Merino dan Ekor Tipis serta Merino dan Ekor Gemuk. Hubungan genetik lintas populasi domba Batur paling dekat dengan Merino dan paling jauh dengan Ekor Gemuk. Individu dalam populasi domba Batur membawa gensitokromb sama dengan Merino dan domba Ekor Tipis. Diduga bangsa Merino dan rumpun domba Ekor Tipis sebagai tetua betina domba Batur.
The morphological characteristic of the Batur sheep are resemble to Merino and hybridization (combination) characteristic of the local and Merino sheep. The genetic status of the Batur sheep not yet known, because there wasn’t exactly pedigree (genealogy) record. The identification of the Batur sheep by morphological characteristic the result wasn’t ucurate. The analysis of the Random Amplyfied Polymorphism DNA (RAPD) genom and Restriction Fragment Length Polymorphism (RFLP) gene of cytochrome b can used to be know similarity of the genetic distance within and between population, genetic status and pedigree of the Batur sheep. Objective of the research are to found the marker of RAPD and RFLP gene of cytochrome b specific for Batur sheep breeds. The RAPD analysis are used genome DNA from 89 samples the Batur , Merino, Garut, Thin-tailed and Fat-tailed whereas the RFLP analysis of cytochrome b gene each are used nine heads respectively. The RAPD analysis are used four type of primer 10 mer and one primer 11 mer, whereas the PCR gene of cytochrome b used two primer type 25 mer. Analysis of the RFLP the cytochrome b gene were used restriction enzyme HaeIII endonucleases. Profile of the monomorphics and polymorphics RAPD and RFLP gen of cytochrome b were identified. Amplification 89 of sheep samples with five RAPD primers type were produced 3921 DNA fragment measuring 100 to 1500 bp. The number of the band monomorphic and polymorphic are each 1946 and 2243 respectively. The analysis of the RFLP of the cytochrome b gene are produced eight fragment type of size 118 to 232 bp. The highest total number of the RAPD bands are sampled sheep Batur (1,382) and lowest Fat-tailed sheep (493). The highest total number of monomorphic bands are sample Batur sheep (729) and lowest Garut sheep (204), while the highest polymorphic band in sample of sheep Batur (653) and lowest are the Merino sheep (90). Band the monomorphic and polymorphic specific to sheep Batur breed is 363 and 1500 bp, Merino 345 and 225 bp, 790 and 530 bp for Garut, Thin-tailed is 615 and 620 bp and the Fat-tailed was 615 and 720 bp. Between individuals in the population Batur and Garut sheep,Thin-tailed and Fat-tailed have lower genetic similarity than individuals in the population Merino. The highest Genetic similarity were between individuals were the population of Merino sheep, and the lowest were Garut sheep, whereas the highest of the cross population were between Batur and Merino sheep and lowest are crosses Thin-tailed and Merino sheep. The closest genetic distance was population of Batur and Merino sheep and the most distant was Merino with Thin-tailed and Merino with Fat-tailed. The genetic relationship of Batur sheep population closest are with Merino and most distant with Fat- tailed. Individuals in the Batur sheep population carry the gene cytochrome b similar with Merino and Thin-tailed sheep. Allegedly Merino and Thintailed breed sheep as female elder (ancestor) of the Batur sheep.
Kata Kunci : Domba, Marka Genetik, RAPD, RFLP Gen Sitokromb