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IDENTIFIKASI IKAN CUPANG (Betta imbelis) TRANSGENIK FOUNDER MEMBAWA GEN PENYANDI HORMON PERTUMBUHAN; Identification of Transgenic Founder Betta Fish (Betta imbelis) Carry Growth Hormone Gene.

Penelitian dilakukan untuk mengidentifikasi keberhasilan introduksi gen penyandi hormon pertumbuhan (Growth Hormone, GH) pada induk F-0 ikan Betta imbellis. Ikan transgenik F-0 dibuat dengan menggunakan metode transfeksi. Identifikasi dilakukan menggunakan metode RT-PCR. RNA total diekstraksi dari embrio pooled sample hasil persilangan induk transgenik dan non-transgenik. Berdasarkan analisis ekspresi gen pada embrio juga menunjukkan adanya aktivitas ekspresi gen GH pada semua perlakuan dibandingkan dengan kontrol (embrio hasil persilangan non-transgenik x non-transgenik). Jumlah individu induk F-0 yang membawa gen GH eksogen berdasarkan analisis PCR dengan DNA template dari sirip ekor adalah sebanyak 16%. Individu positif membawa gen GH eksogen tersebut dibesarkan lebih lanjut untuk memproduksi Betta imbellis transgenik F-1. Kandidat ikan transgenik jantan F-0 dikawinkan dengan ikan non-transgenik betina, sedangkan transgenik F-0 betina dikawinkan dengan non-transgenik jantan. Sebanyak 30-50 butir embrio hasil pemijahan F-0 digabung, kemudian DNA genom diekstrak. Sebagian embrio digunakan untuk ekstraksi RNA total untuk analisis ekspresi mRNA GH eksogen. Hasil analisis PCR menunjukkan bahwa semua sampel embrio dari induk transgenik F-0 dapat terdeteksi gen GH eksogen, sedangkan untuk kontrol (non-transgenik) tidak terdeteksi. Ekspresi mRNA juga terdeteksi pada embrio F-1. Dengan demikian, metode transfeksi embrio Betta imbellis efektif digunakan untuk menghasilkan ikan transgenik, dan sangat berpotensi menghasilkan individu F-1 Betta imbellis dengan pertumbuhan lebih cepat. The study was conducted to identify the successful introduction of the growth hormone gene (Growth Hormone, GH) on the F-0 Betta imbellis broodstock. The F-0 transgenic fish was made through transfection methods. Identification was done using RT-PCR method. Total RNA was extracted from pooled embryos sample. Based on the analysis of gene expression in embryos also showed activity GH gene expression in all treatments compared to the control (non-transgenic x non-transgenic). The number of individuals F-0 which carried exogenous GH gene by PCR analysis of the DNA template of the tail fin was as much as 16%. Positive individuals carried the exogenous GH gene raised further to produce transgenic F-1 B. imbellis. Candidate of transgenic F-0 males fish were mated with non-transgenic female fish, whereas the transgenic F-0 females were mated with non-transgenic males. The 30-50 embryos obtained were combined, then their genomic DNA were extracted. Some of the embryos was used for the extraction of total RNA for analysis of mRNA expression of GH exogenous. The PCR analysis showed that all samples of embryos from the transgenic F-0 broodstock could be detected, whereas for the control (non-transgenic) was not detected. mRNA expression was also detected in embryos of F-1. The average weight of the F-0 broodstocks were 1.55 g and a total length was 12.97 cm. Thus, the transfection methods through betta embryos peaceful effectively generated transgenic fish, and potentially produced fast growth of individuals F-1 Betta imbellis.

STIMULASI PERTUMBUHAN JUVENIL ABALON, Haliotis squamata DENGAN PEMBERIAN HORMON REKOMBINAN IKAN rElGH

Masalah yang paling utama dalam budidaya abalon tropis adalah pertumbuhan yang lambat. Penggunaan rElGH (recombinant giant grouper, Epinephelus lanceolatus growth hormone) untuk menstimulasi pertumbuhan beberapa spesies ikan sudah dilakukan. Penelitian ini bertujuan untuk menguji akselerasi pertumbuhan juvenil abalon tropis, Haliotis squamata setelah diberi perlakuan perendaman hormon rekombinan ikan kerapu kertang, Epinephelus lanceolatus pada frekuensi yang berbeda. Ada empat perlakuan frekuensi perendaman rElGH yaitu 4, 9, 16 kali, dan tanpa perendaman (kontrol). Masing-masing perlakuan diulang tiga kali. Perendaman dilakukan selama tiga jam, dengan interval waktu empat hari. Kepadatan abalon tropis 100 ekor/L air laut yang mengandung 30 mg rElGH. Wadah untuk perendaman berupa beaker glass yang dilengkapi dengan aerasi. Penelitian dilakukan selama tujuh bulan. Hasil penelitian menunjukkan bahwa abalon tropis yang direndam rElGH dengan frekuensi empat kali menghasilkan pertumbuhan bobot tubuh dan panjang cangkang tertinggi dan berbeda nyata dengan perlakuan lainnya (P<0,05). Sintasan abalon tropis yang diberi perlakuan perendaman hormon rElGH lebih tinggi dibandingkan perlakuan kontrol. The most crucial problem in tropical abalone aquaculture is the slow growth of the species. Studies investigating the use of rElGH (recombinant giant grouper, Epinephelus lanceolatus growth hormone) for promoting growth have been performed in various species. This research aimed to examine the growth acceleration of tropical abalone, Haliotis squamata juvenile after being treated in different immersion frequencies of recombinant giant grouper, Epinephelus lanceolatus growth hormone (rElGH). There were four treatments of rElGH immersion frequency: 4, 9, 16 times and without rElGH immersion (control). Each treatment was performed in triplicates. Immersion was performed for 3 hours, at 4-day intervals and a density of 100 tropical abalones in 1 L seawater containing 30 mg rElGH. Immersion was conducted in a beaker glass supplied with oxygen. The results indicated that rElGH immersion for total of 4 times showed significantly higher (P<0.05) body weight and shell length of tropical abalone compared to other treatments. The Survival of tropical abalone treated with rElGH was also significantly higher than control.