Kajian Keragaman genetik Kuskus (Phalangeridae) Asal Maluku dan Papua Berdasarkan Sekuen Gen Penyandi NADH Dehidrogenase Subunit 4

Morsid Andityas, Dr. drh. Rini Widayanti, M.P.; Dr. drh. Hery Wijayanto, M.P.

2016 | Tesis | S2 Sain Veteriner

Kuskus merupakan mamalia marsupialia yang termasuk dalam famili Phalangeridae. Keberadaan satwa ini tersebar di Indonesia bagian timur yaitu Sulawesi, Maluku, Pulau Timor dan Papua. Usaha konservasi untuk melindungi kuskus dari kepunahan masih terhambat dikarenakan data tentang satwa ini yang terbatas. Penelitian ini bertujuan mengidentifikasi sekuen nukleotida dan keragaman genetik gen ND4 pada kuskus asal Maluku dan Papua. Sebanyak 21 sampel didapatkan dari 13 sampel asal Maluku dan 8 sampel asal Papua. Sampel diisolasi DNA totalnya dan diamplifikasi dengan metode PCR. Produk hasil PCR sebesar 1556 base pair kemudian dilakukan sekuensing. Hasil sekuensing dianalisis dengan program MEGA versi 7. Sekuen gen ND4 dilakukan penjajaran berganda dengan spesies kuskus lain dari genbank menggunakan Clustal W. Analisis sekuen asam amino dari basa-basa diterjemahkan mengikuti vertebrate mitochondrial translation code. Produk hasil sekuensing sebesar 1380 nukleotida dari gen ND4 digunakan untuk analisis filogenetik. Berdasarkan konstruksi pohon filogenetik dengan menggunakan metode Neighbor Joining pada sekuen gen ND4 terdiri dari kelompok Spilocuscus dan Phalanger. Terdapat 51 situs nukleotida yang dapat digunakan sebagai penanda genetik antara kelompok Phalanger asal Papua dengan Maluku dan 31 situs nukleotida untuk penanda genetik antara kelompok Spilocuscus asal Papua dengan Maluku. Penanda genetik antara kelompok Spilocuscus asal Papua dengan Pulau sentani dan Tobelo Maluku Utara terdapat 13 situs nukleotida, sedangkan antara kelompok Spilocuscus asal Maluku dengan Pulau Sentani dan Tobelo Maluku Utara terdapat 11 situs nukleotida.

Cuscuses is a marsupial mammals which is a member of family Phalangeridae. This animal's presence is spread out in eastern Indonesia consists of Sulawesi, Maluku, Timor Island and Papua. Conservation effort to protect cuscuses from extinction is still hampered due to lack of information about this animal. This study aims to identify nucleotides sequences and genetic diversity of ND4 gen in kuskus from Maluku and Papua. There are 21 total samples consist of 13 samples from Maluku and 8 samples from Papua. The total DNA was extracted and amplified from sample by PCR. The result of PCR, 1556 base pair were sequenced. The result of sequencing was analyzed using MEGA version 7. ND4 gen sequences was multiple alignment with another cuscuses species from genbank using Clustal W. The amino acid analysis from bases were translated followed vetrebrae mitochondrial translation code. 1380 nucleotides from ND4 gen from sequencing used for philogenetic analysis. Based on philogenetic tree construction by Neighbor Joining method, the ND4 gen sequence showed Spilocuscus and Phalanger group as a result. There were 51 nucleotides site that can be used as genetic marker between Phalanger from Papua and Maluku. Furthermore, there are 31 nucleotides site that can be used as genetic marker for Spilocuscus from Papua and Maluku. There were 13 nucleotides sites that can be used as genetic marker for Spilocuscus sample from Papua compared to sample from Sentani Island and Tobelo North Maluku. There were 11 nucleotides sites that can be used as genetic marker between Spilocuscus sample from Maluku and sample from Sentani Island and Tobelo North Maluku.

Kata Kunci : Kuskus, gen ND4, Sekuensing, Penanda Genetik