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Desain Primer PCR Pendeteksi Gen Imun Non Spesifik Toll-Like Receptor (TLR3) dan Chemocine (CXCL10) pada ikan lele (Clarias gariepinus)

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ABSTRAK

Imunitas merupakan mekanisme utama pertahanan tubuh makhluk hidup yang terdiri dari dua bagian yakni sistem imun non spesifik dan sistem imun spesifik. Imun Toll-Like Receptors (TLR3) diyakini memainkan peranan penting dalam pertahanan tubuh untuk mencegah dan menghambat invasi patogen dalam sistem imun non spesifik. Chemocine (CXCL10) merupakan keluarga protein-protein kecil yang diproduksi oleh jaringan terinfeksi pada tahapan awal infeksi. Penelitian ini bertujuan untuk mendesain primer spesifik yang mampu mengamplifikasi gen TLR3 dan CXCL10 pada spesies ikan lele (C. gariepinus). Primer PCR gen imun tersebut didesain berdasarkan gen imun yang telah dikarakterisasi dan diyakini memiliki gen imun TLR3 dari ikan lele Clarias batrachus dan CXCL10 dari ikan Catfish Ictalurus punctatus dan I. furcatus. Sampel DNA untuk pengujian diambil dari bagian jaringan limfomieloid, yaitu organ limpa, ginjal dan hati yang berperan dalam memproduksi sel-sel darah dan respon imun, baik seluler maupun non spesifik. Primer TLR3 yang didesain mampu mengamplifikasi gen imun TLR3 pada ikan lele (C. gariepinus) sebanyak 699 bp, sedangkan primer CXCL10 didesain secara in silico, namun tidak diamplifikasi karena dianggap kurang memenuhi/sesuai digunakan sebagai primer yang baik, sehingga dikhawatirkan tidak akan mampu mengamplifikasi gen imun CXCL10 pada ikan lele (C. gariepinus). Hasil PCR gen imun TLR3 ikan lele (C. gariepinus) kemudian disekuensing, hasil sekuensing kemudian dihomologikan dengan gen imun TLR3 yang terdapat di Gene Bank. Hasil homologi gen TLR3 tersebut memiliki homologi dengan gen Imun TLR3 ikan Lele (Clarias batrachus) momor aksesei KC. 907862 sebanyak 96%.


Kata kunci : Clarias gariepinus, Ictalurus punctatus dan Ictalurus furcatus, Toll-Like receptors 3 (TLR3), Chemocine (CXCL10), desain primer

ABSTRACT

Immunity is the main defence mechanism of living organism, composed of two parts namely non-specific immune system and specific immune system. Toll-Like Receptors (TLR3) was believed playing an important role in the body's defenses to prevent and inhibit pathogens invation in non-specific immune system and Chemocine is a family of a small proteins that produced by the infected tissue in the early stages of infection. This research aims to design specific PCR primers that capable of amplifying the TLR3 and CXCL10 gene’s on Catfish species (Clarias gariepinus). PCR primers were designed based on the immune genes that have been characterized and believed has TLR3 in catfish species C. batrachus and CXCL10 immune gene from Ictalurus punctatus and I. furcatus. DNA samples for the experiment were taken from the tissue of limfomieloid organ. The lymphomyeloid organs were spleen, kidneys and liver that plays a role in producing blood-cells and the immune response both cellular and humoral. Primer TLR3 was designed and could amplify the TLR3 gene on local Catfish (C. gariepinus) and produce approximately 699 bp. CXCL10 primer was designed in silico, but not be amplified because they did not meet/suitable for use as a good primer, so it worried not be able to amplify CXCL10 immune genes on catfish species (C.gariepinus). The product of PCR of TLR3 gene from catfish (C. gariepinus) was sequenced. Furthermore, the sequence was analized similiarity comparison with the exsist TLR3 immune genes found in Gene Banks collections. The result showed a similiarity comparison between TLR3 gene from C. gariepinus with TLR3 gene from Catfish species (Clarias batrachus) accession number KC. 907862 as much as 96%.

Keyword : Clarias gariepinus, Ictalurus punctatus and Ictalurus furcatus, Toll-Like receptors 3 (TLR3), Chemocine (CXCL10), primer design.

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Ketersediaan

1108B17II

576 FRA d

Perpustakaan FSM Undip (Referensi)

Tersedia

Informasi Detail

Judul Seri

BIOLOGI

No. Panggil

576 FRA d

Penerbit

: ., 2017

Deskripsi Fisik

-

Bahasa

Indonesia

ISBN/ISSN

-

Klasifikasi

1588

Tipe Isi

-

Tipe Media

-

Tipe Pembawa

-

Edisi

-

Subjek

-

Info Detail Spesifik

-

Pernyataan Tanggungjawab

Frans Oktavianus Siregar

Versi lain/terkait

Lampiran Berkas

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