ISOLASI DNA PARSIAL GEN Tyrosinase-Related Protein-1 (TYRP1) PADA IKAN GURAME (Osphronemus gouramy)

• Main Author: Sandy, Angga Kly (Author)
• Format: Book
• Published: 2015-11-25.
• Subjects: Thesis; NonPeerReviewed
• Online Access: Link Metadata
• Notes: http://repository.upi.edu/21836/1/S_BIO_1105533_Title.pdf; http://repository.upi.edu/21836/2/S_BIO_1105533_Abstract.pdf; http://repository.upi.edu/21836/3/S_BIO_1105533_Table_of_content.pdf; http://repository.upi.edu/21836/4/S_BIO_1105533_Chapter1.pdf; http://repository.upi.edu/21836/5/S_BIO_1105533_Chapter2.pdf; http://repository.upi.edu/21836/6/S_BIO_1105533_Chapter3.pdf; http://repository.upi.edu/21836/7/S_BIO_1105533_Chapter4.pdf; http://repository.upi.edu/21836/8/S_BIO_1105533_Chapter5.pdf; http://repository.upi.edu/21836/9/S_BIO_1105533_Bibliography.pdf; http://repository.upi.edu/21836/10/S_BIO_1105533_Appendix1.pdf; http://repository.upi.edu/21836/11/S_BIO_1105533_Appendix2.pdf
• Keywords: QH426 Genetics; QL Zoology

Tyrosinase-Related Protein-1 (TYRP1) merupakan salah satu gen yang berperan dalam proses sintesis melanin. Gen TYRP1 memiliki peranan penting dalam menginstruksikan pembentukan enzim tyrosinase-related protein-1. Ekspresi gen TYRP1 dapat digunakan sebagai penanda kondisi stres pada ikan yang dapat disebabkan oleh faktor internal dan eskternal. Namun informasi genetik mengenai gen TYRP1 pada ikan gurame (Osphronemus gouramy) sangat minim informasi, sehingga pada penelitian ini bermaksud untuk mengisolasi dan mengarakterisasi gen TYRP1 pada ikan gurame. Amplifikasi gen TYRP1 pada DNA genom ikan gurame di lakukan dengan menggunakan dua pasang primer degenerate yang di rancang berdasarkan konsensus sikuen gen TYRP1 dari beberapa ikan dari infraclass Teleostei. Perancangan primer dilakukan secara online pada laman Primaclade dan Beacon Designer Free Edition. Primer terdiri dari set outer-primer dan inner-primer (nested), dimana outer-primer digunakan untuk memulai proses amplifikasi, kemudian produk amplifikasi yang tidak diharapkan dapat diminimalisir dengan keberadaan inner-primer. Analisa hasil sequencing dilakukan dengan BLAST, dan didapat hasil homologi pada ikan Takifugu rubripes (AF397401.1) dengan nilai kesamaan (Ident.) sebesar 76%. Sikuen DNA parsial gen TYRP1 ikan gurame dengan panjang 525 pb yang terdiri dari 36 pb daerah intron dan 489 pb daerah exon didapatkan dari hasil pensejajaran sikuen sampel dengan outer-primer dengan sikuen sampel dengan inner-primer menggunakan software Bioedit versi 7.2.5. Gen TYRP1 berhasil teramplifikasi dari DNA genom ikan gurame menggunakan dua pasang primer degenerate yang di rancang berdasarkan sikuen sekerabat ikan gurame dari infraclass Teleostei. Di lakukan perancangan primer spesifik gen TYRP1 ikan gurame dari sikuen daerah exon gen TYRP1 ikan gurame yang telah didapatkan. ---------- The Tyrosinase-Related Protein-1 (TYRP1) gene provides instructions for making an enzyme called tyrosinase-related protein-1 in the melanin biosynthesis pathway. TYRP1 enzyme function is to assist in the process of melanogenesis or synthesis melanin. TYRP1 gene expression can be used as a sign that the fish (and other vetebrates) under stress condition, either due to environmental disturbances or diseases. However the genetic information of the TYRP1 genes doesn't exist in goramy (Osphronemus gouramy) or minimum shared information. In this study focus to isolate and characterize the TYRP1 genes on gouramy. Amplification of TYRP1 genes in the gouramy DNA genome performed using degenerate primers were designed based on sequence consensus TYRP1 genes of some Teleostei fish. Primer designed by Primaclade and to determining the secondary structure of primer using Beacon Designer Free Edition. First, used outer-primer for initial phase of amplification, and then non specifics products can be eliminated by nested-primer (inner-primer). Four sequences based on the results of sequencing performed contig and then obtained length 525 bp of DNA genom region. Intron and exon are exist in sequence. The identities percentage of sequence is 76 % with Takifugu rubripes (AF397401.1) in BLAST result. Gene sequences analysis obtained TYRP1 goramy exon region with sequence length 489 bp and 36 bp intron region. TYRP1 gene successfully amplified from the DNA genome of goramy using degenerate primers were designed based on sequences of Telesotei infraclass. Design of TYRP1 gene spesific primer on goramy was performed using exon region sequence.