Kusumawati, Daniar (2011) Kajian Gen Pengkode Pola Pigmen dan Profil Protein pada Ikan badut Hitam (Amphiprion percula). Magister thesis, Universitas Brawijaya.
Abstract
Dalam perkembangan budidaya ikan badut hitam ( Amphiprion percula ) sebagai upaya konservasi menemui kendala yaitu munculnya degenaratif pola pigmen pada populasi hasil budidaya jika dibandingkan dengan populasi alam. Karakter fenotip pola pigmen merupakan salah satu karakter yang dikontrol secara genetik. Diketahui bahwa terdapat 94 gen dan 285 alel yang berperan dalam perkembangan normal warna dan pola pigmen pada ikan hias zebra Danio rerio , tiga diantaranya adalah gen tyrosinase, Mitf, dan endothelin receptor b1. Tujuan dari penelitian ini adalah untuk mengkaji peran gen tyrosinase, Mitf, dan endothelin receptor b1 dalam mengkode pola pigmen ikan Badut hitam ( Amphiprion percula ) serta peran lingkungan terhadap profil protein berdasarkan karakter fenotip. Penelitian ini dilaksanakan di Balai Besar Perikanan Budidaya Laut Gondol Propinsi Bali dan di Laboratorium Biologi Molekuler Fakultas MIPA Universitas Brawijaya, Malang. Sampel yang digunakan dalam penelitian ini adalah populasi alam yang diwakili oleh koleksi induk-induk ikan Badut dari habitat alami dan populasi budidaya yang diwakili oleh koleksi benih-benih hasil budidaya. Masing-masing sampel diseleksi berdasarkan karakter fenotip yang muncul kemudian di lakukan ekstrasi DNA dengan menggunakan QIAamp DNA Mini Kit Qiagen (50). Kemudian analisis gen pengkode warna tyrosinase, Mitf dan endothelin receptor b1 (ednrb1) dilakukan dengan mengamplifikasi genom ikan badut hitam menggunakan primer tyr (tyrosinase), nac (mitf), dan rose (ednrb1) melalui program speedy PCR. Selanjutnya dilakukan analisis SSCP (single strand confirmation polymorphism) untuk mengidentifikasi variasi genetic dari gen-gen pengkode warna (tyrosinase, Mitf, endothelin receptor b1). Sedangkan untuk mengetahui peran lingkungan terhadap profil protein pada organ kulit yang dilakukan dengan ekstraksi protein menggunakan metode eletroforesis SDS PAGE. Sampel yang digunakan adalah sampel populasi budidaya yang dipelihara dalam dua kondisi lingkungan berbeda yaitu indoor dan outdoor. Berdasarkan hasil penelitian menunjukkan bahwa primer rose (ednrb1) belum dapat mengamplifikasi genom DNA ikan badut hitam. Sedangkan primer tyr (tyrosinase) dan nac (mitf) dapat mengamplifikasi genom DNA ikan badut hitam pada target sequence 70 bp dan 200 bp. Berdasarkan pola pita fragmentasi amplicon prime tyr dan nac hasil analisis SSCP menunjukkan ketiadaan korelasi dengan pola degeneratif pola pigmen pada ikan badut hitam. Pada eksperimen pemeliharaan dengan dua kondisi yang berbeda yaitu outdoor dan indoor menunjukkan adanya korelasi positif antar profil pita protein kulit dengan pola pigmen ikan badut hitam. Diketahui bahwa intensitas cahaya merupakan faktor eksternal yang diketahui memberikan korelasi positif terhadap migrasi pola pigmen.
English Abstract
In the development of culture of marine ornamental fish Amphiprion percula for the conservative effort, it encounters many challenges. One of this obstacles is population of cultured black clownfish shows degenerative of pigment pattern from that of the wild type. Phenotipic character has been known as a one of genetically controlled character. Approximately 90 genes and 285 alels have been identified in zebrafish that play role in a normal development of colour and pigment pattern. Three of those genes are tyrosinase, Mitf, and endothelin receptor b1. The aim of this experiment are to evaluate the role of tyrosinase, Mitf and endothelin receptor b1 genes that encode pigment pattern of black clown fish ( Amphiprion percula and to evaluate the role of environment towards proteins profiles based on phenotypic character. This research was conducted in The Research Institute of Mariculture, Gondol, Bali and Laboratory of Biochemistry, Faculty of Mathematics and Science Education, Brawijaya University, Malang. Sample used in this experiment were populations from culture which is representative of F1 progeny of culture, and population from nature which is representative of broodstock. Each sample was selected based on phenotypic character, then extraction of DNA carried out by using QIAamp DNA Mini Kit (50) (USA). Afterward, analysis of gene encode pigment pattern was amplified with three primer (rose, tyr, nac) by using speedy PCR program. Then, SSCP analysis was conducted to identify genetic variation of product PCR, followed by sequencing on band profile which is shows difference between cultural population and natural population samples. Meanwhile, the isolation of protein by SDS PAGE method was carried out to determine the extent of the role of the environment affect the protein profile. Sample used in the analysis of protein were population of culture that placed in two different conditions indoor and outdoor. Based on this research, the primary rose (ednrb1) has not amplified the DNA genom of black clown fish. In another, the primary of tyr (tyrosinase) and nac (mitf) has amplified the DNA genome of this fish on sequence 70 bp and 200 bp. Based on the fragmentation amplicon primer tyr pattern of bands and nac from SSCP analysis shown that no correlation with the pattern degenerative of pigment pattern in black clown fish. In the rearing experiment with two different condition which outdoor and indoor were shown the positive correlation between bands of skin protein profile with the pigment pattern of black clown fish. The light intensity has been known as an external factor that gives a positive correlation with the migration of pigment pattern.
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/572.59/KUS/k/041103237 |
| Subjects: | 500 Natural sciences and mathematics > 572 Biochemistry > 572.5 Miscellaneous chemicals |
| Divisions: | S2/S3 > Magister Biologi, Fakultas MIPA |
| Depositing User: | Endro Setyobudi |
| Date Deposited: | 30 Sep 2011 16:45 |
| Last Modified: | 30 Sep 2011 16:45 |
| URI: | http://repository.ub.ac.id/id/eprint/157677 |
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