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All Journal HAYATI Journal of Biosciences Jurnal Akuakultur Indonesia Agromet Jurnal Ilmu dan Teknologi Kelautan Tropis Proceedings of Annual International Conference Syiah Kuala University - Life Sciences & Engineering Chapter BIOTROPIA - The Southeast Asian Journal of Tropical Biology Indonesian Journal of Biotechnology Widyariset KOVALEN: Jurnal Riset Kimia Omni-Akuatika Jurnal Perikanan Universitas Gadjah Mada Kelola: Journal of Islamic Education Management Indonesian Aquaculture Journal Jurnal Riset Akuakultur Journal of Educational Research and Evaluation BJRA (Bongaya Journal of Research in Accounting) Efektor JSMA (Jurnal Sains Manajemen dan Akuntansi) Widyariset GANDRUNG: Jurnal Pengabdian Kepada Masyarakat Siyasatuna Jurnal Ilmiyah Mahasiswa Siyasah Syariyyah Jurnal Aktiva : Riset Akutansi dan Keuangan Qadauna: Jurnal Ilmiah Mahasiswa Hukum Keluarga Islam Journal of Applied Civil Engineering and Infrastructure Technology (JACEIT) Jumper : Jurnal Mahasiswa Pendidikan Olahraga Jurnal Media Akuakultur Indonesia Shautuna: Jurnal Ilmiah Mahasiswa Perbandingan Mazhab Arus Jurnal Sosial dan Humaniora Jurnal Pengabdian Inovasi dan Teknologi Kepada Masyarakat SinarFe7 Private Law Journal of Applied Sciences in Accounting, Finance, and Tax Jurnal Ilmiah Pendidikan Dasar (JIPDAS) Jurnal Kedokteran Hewan Jurnal Pengabdian Masyarakat Olahraga dan Kesehatan Vokatek : Jurnal Pengabdian Masyarakat PEDAMATH: Journal on Pedagogical Mathematics Al Fuadiy: Jurnal Hukum Keluarga Islam Economos : Jurnal Ekonomi dan Bisnis Journal Sultra Research of Law International Journal of Educational and Life Sciences (IJELS) Jurnal Media Akademik (JMA) ENLEKTURER: Journal of Islamic Studies Akrual: Jurnal Bisnis dan Akuntansi Kontemporer HISABUNA: Jurnal Ilmu Falak Journal of Fisheries & Marine Inovasi Teknologi Masyarakat Journal of Mathematics, Computation and Statistics (JMATHCOS) International Journal of Education, Vocational and Social Science Hikamatzu Journal of Multidisciplinary Research Jurnal Bioteknologi & Biosains Indonesia (JBBI) Bersama: Jurnal Pengabdian Kepada Masyarakat Issues in Mathematics Educations (IMED) Journal of Pedagogical and Teacher Professional Development Lumbung Ngabdi: Jurnal Pengabdian Masyarakat At-Tadris: Journal of Islamic Education J-Sil (Jurnal Teknik Sipil dan Lingkungan) Jurnal Bioteknologi & Biosains Indonesia Journal of Arts Education and Design

Alimuddin

Departemen Budidaya Perairan, Fakultas Perikanan Dan Kelautan, Institut Pertanian Bogor Jl. Agatis, Kampus IPB Dramaga 16680

Author-ID : 2471926

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Aplication of Gene Transfer in Aquaculture Alimuddin, .; Yoshizaki, G.; Carman, O.; Sumantadinata, K.
Jurnal Akuakultur Indonesia Vol. 2 No. 1 (2003): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Recently, global food security has become a hot issue by the public in national as well as out of the country. Aquacultural output will need to be increased several fold in order to meet the rising demands for fish in coming years as the increasing of mankind population. The intensity and capacity of production is expected to increase using biotechnology approach. One of the advances biotechnologies that expected to be a powerful approach for aquaculture development is transgenic technique. This technique has been applied to several commercially valuable species. This review describes various techniques of gene transfer, persistence and expression of transferred gene, application and future aspect of gene transfer research in aquaculture. Key words: Gene transfer, gene expression, biotechnology, aquaculture ABSTRAK Saat ini, keamanan pangan telah menjadi isu hangat di masyarakat baik di dalam maupun di luar negeri. Produksi akuakultur diharapkan dapat ditingkatkan beberapa kali lipat untuk memenuhi kebutuhan pangan berupa ikan dimasa-masa mendatang akibat peningkatan populasi manusia. Intensitas dan kapasitas produksi diharapkan meningkat dengan menggunakan pendekatan bioteknologi. Salah satu teknik modern yang diduga akan menjadi sarana yang berguna dalam pengembangan akuakultur adalah teknologi transfer gen. Teknik ini telah diaplikasikan pada spesies-spesies yang memiliki nilai ekonomis. Ulasan ini menggambarkan variasi metode transfer gen, persistensi dan ekspressi dari gen yang ditransfer, aplikasi dan prospeknya ke depan dari penelitian transfer gen dalam akuakultur. Kata kunci : transfer gen, ekspressi gen, bioteknologi, akuakultur

Effect of Temperature on Reproduction and Sex Ratio of Guppy (Poecilia reticulata Peters) Arfah, H.; Mariam, S.; Alimuddin, .
Jurnal Akuakultur Indonesia Vol. 4 No. 1 (2005): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Water temperature could affect the reproduction of broodstock and sex ratio of progeny. In this study, broodstock of guppy (Poecilia reticulata Peters) was reared in different temperature to determine its effect on reproduction of broodstock and sex ratio of their progeny. The result of study show that broodstock reared at 27°C produced more fry (16 males mean) than that of 30°C (10 males), while broodstock reared at 33°C produced no progeny. Percentage of male fish produced by broodstock reared at 30°C is higher than that of 27°C. Incubation time of embryo before birth is sorter in broodstock reared at 30°C (4-12 days) compared with 27°C (18-22 days). However, several progeny of broodstock reared at 30°C had abnormal vertebrae. Keywords: guppy, Poecilia reticulata, sex reversal, reproduction, monosex ABSTRAK Suhu air inkubasi diduga dapat mempengaruhi reproduksi induk ikan dan nisbah kelamin keturunannya. Pada penelitian ini, induk ikan gapi (Poecilia reticulata Peters) dipelihara pada suhu 27°C, 30°C dan suhu 33°C untuk mengetahui pengaruhnya terhadap reproduksi dan rasio kelamin keturunannya. Hasil penelitian menunjukkan bahwa induk ikan gapi yang dipelihara pada suhu 27°C menghasilkan anak lebih banyak (rata-rata 16 ekor) daripada di suhu 30°C (10 ekor), sementara induk gagal melahirkan pada suhu pemeliharaan 33°C. Proporsi anak jantan yang dihasilkan oleh induk yang dipelihara pada suhu 30°C lebih banyak dibandingkan pada suhu 27°C. Waktu inkubasi embrio sebelum dilahirkan oleh induk yang dipelihara pada suhu 30°C lebih singkat, yaitu 4-12 hari, dibandingkan pada suhu 27°C, 18-22 hari. Namun demikian beberapa anak ikan yang lahir dari induk yang dipelihara pada suhu suhu 30°C mengalami abnormalitas pada bagian tulang belakangnya. Kata kunci: ikan gapi, Poecilia reticulata, sex reversal, reproduksi, monoseks

Effectiveness of hCMV, mEF1a and mAct promoters on driving of foreign gene expression in transgenic zebrafish Alimuddin, .; Yoshizaki, G.; Carman, O.; Takeuchi, T.
Jurnal Akuakultur Indonesia Vol. 6 No. 1 (2007): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Highly unsaturated fatty acids (HUFA), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have long been recognized for its beneficial effect for human health and development. The D6 fatty acid desaturase is generally considered to be the rate-limiting factor in HUFA biosynthesis. Here, as the first step of study, we conducted experiment to select an appropriate construct that allows higher expression levels of masu salmon (Oncorhynchus masou) D6-desaturase gene in zebrafish (Danio rerio) in order to increase its activity for synthesizing EPA/DHA. Salmon D6-desaturase cDNA (sD6) was separately ligated with human cytomegalovirus (hCMV), medaka elongation factor 1a (mEF1a) and medaka b-actin (mAct) promoters. The resulted construct was designated as hCMV-sD6, mEF1a-sD6 and mAct-sD6, respectively. Each of the constructs in circular DNA form was microinjected into 1-cell stage embryos at a concentration of 30mg/ml. Transgenic individuals were identified by polymerase chain reaction (PCR) and their expression levels were analyzed by reverse transcription PCR. The first (F1) and second (F2) generation was produced by crossing the transgenic founder F0 and F1, respectively, with wild-type fish. The results showed that the highest transient gene expression level was obtained from the mAct-D6 construct, followed respectively by EF1a-D6 and hCMV-D6 construct. The transmission rate of transgene into F1 generation was 4.2%-44.1%, and into F2 was followed the Mendellian segregation pattern. Expression of transgene in F2 generation was varied between strains regarding as the mosaics of F0 fish. Now, a transgenic system to study the modification of fatty acid biosynthesis pathways in fish was established. Further investigations are to produce fish containing higher levels of EPA and DHA. Keywords: desaturase, nutraceutical fatty acid, transgenic, zebrafish, masu salmon Abstrak Promoter merupakan regulator yang menentukan tempat, waktu dan tingkat ekspresi gen. Pada penelitian ini, kami melakukan seleksi kontruksi plasmid yang tepat yang menghasilkan tingkat ekspresi yang tinggi dari gen D6-desaturase-like ikan masu salmon (Oncorhynchus masou) yang ditransfer ke ikan zebra (Danio rerio) untuk meningkatkan kemampuannya dalam mensintesa EPA/DHA. cDNA D6-desaturase-like (OmD6FAD) dari ikan salmon masu diligasi secara terpisah dengan promoter dari cytomegalovirus manusia (hCMV), elongation factor 1a (mEF1a) dan b-actin (mbAct) dari ikan medaka, untuk membuat konstruksi plasmid yang berturut-turut disebut sebagai hCMV-OmD6FAD, mEF1a- OmD6FAD dan mbAct-OmD6FAD. Konstruksi tersebut dengan konsentrasi 30mg/ml disuntikkan ke embrio pada saat fase satu sel. Individu transgenik diidentifikasi menggunakan PCR dan tingkat ekspresi transgen dianalisa dengan RT-PCR. Hasil menunjukkan bahwa tingkat ekspresi sementara yang tertinggi dari gen asing adalah diperoleh dari konstruksi mbAct-OmD6FAD, diikuti selanjutnya oleh EF1a-OmD6FAD dan hCMV- OmD6FAD. Transgen telah ditransmisikan ke ikan generasi F2 dengan mengikuti pola segregasi Mendel. Tingkat ekspresi transgen yang tinggi pada jaringan ikan F2 yang diperiksa telah diperoleh. Dengan demikian, sebuah sistem transgenik untuk memodifikasi biosistesa asam lemak pada ikan telah dikembangkan. Kata kunci: promoter, desaturase asam lemak, transgenik, ikan zebra, ikan salmon masu

Rapid method for identification of transgenic fish zygosity Alimuddin, .; Yoshizaki, G.; Carman, O.
Jurnal Akuakultur Indonesia Vol. 6 No. 2 (2007): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Identification of zygosity in transgenik fish is normally achieved by PCR analysis with genomic DNA template extracted from the tissue of progenies which are derived by mating the transgenic fish and wild-type counterpart. This method needs relatively large amounts of fish material and is time- and labor-intensive. New approaches addressing this problem could be of great help for fish biotechnologists. In this experiment, we applied a quantitative real-time PCR (qr-PCR) method to analyze zygosity in a stable line of transgenic zebrafish (Danio rerio) carrying masu salmon, Oncorhynchus masou D6-desaturase-like gene. The qr-PCR was performed using iQ SYBR Green Supermix in the iCycler iQ Real-time PCR Detection System (Bio-Rad Laboratories, USA). Data were analyzed using the comparative cycle threshold method. The results demonstrated a clear-cut identification of all transgenic fish (n=20) classified as a homozygous or heterozygous. Mating of those fish with wild-type had revealed transgene transmission to the offspring following expected Mendelian laws. Thus, we found that the qTR-PCR to be effective for a rapid and precise determination of zygosity in transgenic fish. This technique could be useful in the establishment of breeding programs for mass transgenic fish production and in experiments in which zygosity effect could have a functional impact. Keywords: quantitative real-time PCR; zygosity; transgenic fish; mass production ABSTRAK Identifikasi sigositas ikan transgenik biasanya dilakukan menggunakan analisa PCR dengan cetakan DNA genomik yang diekstraksi dari jaringan ikan hasil persilangan antara ikan transgenik dan ikan normal. Metode ini memerlukan ikan dalam jumlah yang banyak, dan juga waktu serta tenaga. Pendekatan baru untuk mengatasi masalah tersebut akan memberikan manfaat besar kepada peneliti bioteknologi perikanan. Pada penelitian ini, kami menggunakan metode PCR real-time kuantitatif (krt-PCR) untuk menganalisa sigositas pada satu strain ikan zebra (Danio rerio) transgenik yang membawa gen D6-desaturase-like dari ikan salmon masu, Oncorhynchus masou. krt-PCR dilakukan menggunakan iQ SYBR Green Supermix pada mesin iCycler iQ Real-time PCR Detection system (Bio-Rad Laboratories, USA). Data dianalisis menggunakan metode pembandingan nilai cycle threshold. Hasil penelitian menunjukkan bahwa semua ikan transgenik (n=20) yang diidentifikasi dapat diklasifikasikan secara jelas sebagai ikan homosigot atau heterosigot. Persilangan antara ikan transgenik tersebut dengan ikan normal menunjukkan transmisi transgen ke keturunannya mengikuti hukum segregasi Mendel. Dengan demikian, metode krt-PCR adalah efektif untuk penentuan sigositas secara cepat dan tepat pada ikan transgenik. Teknik ini dapat berguna dalam program produksi ikan transgenik secara massal dan dalam percobaan dimana faktor sigositas memberikan pengaruh nyata. Kata kunci: kuantitatif real-time PCR; sigositas, ikan transgenik; produksi massal

Oral Administration of 17α-Methyltestosterone Increased Male Percentage of Freshwater Crayfish Cherax quadricarinatus Carman, O.; Jamal, M.Y.; Alimuddin, .
Jurnal Akuakultur Indonesia Vol. 7 No. 1 (2008): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Cherax quadricarinatus is one of freshwater crayfish species that has enormous potential for expanding its farming in future. Application of monosex male culture using steroid sex hormone administration method during the period of sex differentiation or early developmental stage might be increased efficiency in farming. This study was aimed to increase male of C. quadricarinatus by oral administration of diet containing 17α-methyltestosterone (MT) towards production efficiency. Two-week-old of Cherax quadricarinatus were fed ad libitum on diets containing various dose of MT, i.e., 25, 50, 75, 100 and 150 mg/kg diet or diet containing no MT as control, 3 times daily for 30 days. After MT-treatment, crayfish were fed frozen Chironomus sp. and shrimp diet. Sex ratio, survival and growth rate (by length and weight) were observed at the end of experiment. Sex was determined by visual observation; the male sex organ is located at the fifth walking leg while the female is at the third. Data was analyzed by F and BNT tests. The results of study show that administration of MT was significantly changed the male ratio of crayfish. Treatment dose of 50 mg/kg diet was effective to increase male sex percentage from 24.93% (control) to be 59.96%. Growth was also significantly being improved, while survival rate was insignificant. Thus, oral administration of MT is an effective way to increase male sex percentage of crayfish, although other methods and the time of hormone administration are needed to be verified to obtain maximal results. Keywords: monosex, 17α-methyltestosterone, sex reversal, Cherax quadricarinatus ABSTRAK Salah satu jenis lobster air tawar yang berpotensi tinggi untuk dikembangkan usaha budidayanya adalah Cherax quadricarinatus. Aplikasi teknik budidaya tunggal kelamin (monoseks) dengan metode pemberian hormon seks steroid yang diberikan pada saat diferensiasi kelamin atau masa perkembangan awal ikan diduga dapat meningkatkan efisiensi usaha. Penelitian ini ditujukan untuk meningkatkan persentase C. quadricarinatus jantan menggunakan metode seks reversal melalui pemberian pakan yang mengandung 17α-metiltestosteron (MT) sebagai upaya efisiensi produksi. C. quadricarinatus umur 2 minggu diberi pakan yang mengandung MT dengan dosis 25, 50, 75, 100 dan 150 mg/kg pakan atau tanpa hormon secara ad libitum, 3 kali sehari selama 30 hari. Setelah perlakuan lobster uji diberi pakan alami Chironomus sp. beku dan pakan udang. Parameter yang diamati meliputi nisbah kelamin, kelangsungan hidup dan pertumbuhannya (panjang dan berat mutlak), yang dilakukan pada akhir penelitian. Identifikasi jenis kelamin dilakukan secara visual; alat kelamin lobster jantan terdapat pada bagian pangkal kaki jalan kelima, yang betina terletak pada bagian dasar kaki jalan ketiga. Data dianalisis menggunakan uji F dan BNT. Hasil penelitian menunjukkan bahwa pemberian hormon MT berpengaruh nyata terhadap persentase kelamin jantan lobster. Perlakuan dengan dosis 50 mg/kg pakan efektif untuk meningkatkan persentase jantan C. quadricarinatus dari 24,93% (kontrol) menjadi 59,96%. Pertumbuhan panjang dan berat mutlak juga menunjukkan pengaruh yang berbeda nyata, sementara kelangsungan hidup tidak berbeda. Dengan demikian pemberian hormon MT melalui pakan cukup efektif untuk meningkatkan persentase lobster jantan yang dihasilkan, meskipun penggunaan metode lain dan waktu pemberian hormon MT masih perlu diteliti untuk memperoleh hasil maksimal. Kata kunci: tunggal kelamin, 17α-metiltestosteron, seks reversal, Cherax quadricarinatus

Characterization of β-Actin Promoter from Nile Tilapia (Oreochromis niloticus) Alimuddin, .; Octavera, A.; Arifin, O.Z.; Sumantadinata, K.
Jurnal Akuakultur Indonesia Vol. 7 No. 2 (2008): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Promoter is one of the factors determining the successful of transgenesis. In this study we isolated and characterized β-actin promoter from Nile tilapia (tiBP) towards production of autotransgenic tilapia. β-actin promoter has high activity in muscle. Sequence of tiBP promoter was isolated by using PCR method. Sequencing was performed using ABI PRISM 3100 machine. Analysis of sequences was conducted using GENETYX version 7 and TFBind softwares. DNA fragment of PCR amplification product digested from the vector cloning was then ligated with pEGFP-N1 to generate ptiBP-EGFP construct. The construct was microinjected into one-cell stage of zebrafish (Danio rerio) embryos to test the tiBP promoter activity. EGFP gene expression was observed by fluorescence microscope. The result of sequence analysis showed that the length of DNA fragment obtained is about 1.5 kb and containing the evolutionary conserved sequences of transcription factor for β-actin promoter including CCAAT, CArG and TATA boxes. Furthermore, tiBP sequence in ptiBP-EGFP construct could regulated GFP expression in muscle of zebrafish embryos injected with the construct. The results suggested that PCR amplification product is the regulator sequence of tilapia β-actin gene. Autotransgenic tilapia can be then produced by changing GFP gene fragment of ptiBP-EGFP construct with genes from tilapia encoding important traits in aquaculture. Keywords: cloning, β-actin promoter, autotransgenic, EGFP, Oreochromis niloticus, Danio rerio ABSTRAK Promoter merupakan salah satu faktor penentu keberhasilan transgenesis. Pada penelitian ini kami mengisolasi dan mengkarakterisasi promoter β-actin dari ikan nila (tiBP) dalam rangka pembuatan ikan nila autotransgenik. Promoter β-actin memiliki aktivitas tinggi pada jaringan otot. Sekuens promoter tiBP diisolasi menggunakan metode PCR. Sekuensing dilakukan menggunakan mesin ABI PRISM 3100. Analisa sekuens menggunakan software GENETYX versi 7 dan TFBind. Fragment DNA hasil amplifikasi PCR yang didigesti dari vektor kloning selanjutnya diligasi dengan pEGFP-N1 untuk membuat konstruksi ptiBP-EGFP. Konstruksi ptiBP-EGFP dimikroinjeksi ke embrio ikan zebra (Danio rerio) fase 1 sel untuk menguji aktivitas promoter tiBP. Ekspresi gen EGFP diamati menggunakan mikroskop fluoresens. Analisa sekuens menunjukkan bahwa panjang fragmen DNA hasil amplifikasi PCR sekitar 1,5 kb dan memiliki faktor transkripsi yang konserf untuk promoter β-actin, yaitu CCAAT, boks CArG dan TATA. Selanjutnya, sekuens tiBP dalam konstruksi ptiBP-EGFP mampu mengendalikan ekspresi gen EGFP pada jaringan otot embrio ikan zebra yang dimikroinjeksi dengan konstruksi tersebut. Dengan demikian dapat disimpulkan bahwa fragmen DNA hasil amplifikasi PCR tersebut merupakan sekuens promoter β-actin ikan nila. Pembuatan ikan nila autotransgenik selanjutnya dapat dilakukan dengan mengganti gen EGFP pada pktBA-EGFP dengan gen-gen asal ikan nila yang mengkodekan karakter penting dalam budidaya ikan. Kata kunci: kloning, promoter β-actin, autotransgenik, EGFP, Oreochromis niloticus, Danio rerio

Hematology of common carp following DNA vaccination and koi herpesvirus challenge test Nuryati, Sri; Maswan, N.A.; Alimuddin, .; Sukenda, .; Sumantadinata, K.; Pasaribu, F.H.; Soejoedono, R.D.; Santika, A.
Jurnal Akuakultur Indonesia Vol. 9 No. 1 (2010): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

The study was aimed to determine the effectiveness of DNA vaccine doses on hematological aspect which represent immune response and its influence on common carp survival rate. DNA vaccines encoding the viral glycoprotein of koi herpesvirus (KHV) have been proved to highly protect the fish under laboratory condition. A dose of 12.5 µg/100 µl vaccine had resulted in a survival rate of 96.67 % during 30 days after challenge test with a lethal dose of KHV. Fish vaccinated using lower doses, i.e. 2.5 and 7.5 µg/100µl showed 100% mortality after 15 and 19 days challenge test respectively, whereas non vaccinated fish as a control showed 100% mortality after 17 days challenge test. Total leucocytes of the vaccinated fish were higher than control until 42 days post vaccination, but declined afterward. Phagocytic index of the vaccinated fish using 12.5 µg/100 µl was declined after 49 days post vaccination or 7 days post challenge test. Key words: DNA vaccine, Koi herpesvirus (KHV), leucocyte, phagocytic index, Cyprinus carpio ABSTRAK Penelitian ini bertujuan untuk menguji pengaruh vaksinasi menggunakan vaksin DNA dengan dosis berbeda terhadap gambaran darah ikan sebagai respresentasi tanggap kebal ikan mas serta pengaruhnya terhadap tingkat kelangsungan hidup ikan mas. Vaksin DNA penyandi glikoprotein koi herpesvirus (KHV) dapat memberikan proteksi yang tinggi pada percobaan skala laboratorium. Vaksinasi dengan dosis 12,5 µg/100µl dapat mempertahankan kelangsungan hidup sebesar 96,67% selama satu bulan setelah uji tantang dengan virus KHV menggunakan dosis letal. Ikan yang divaksin dengan dosis yang lebih rendah yaitu 2,5 dan 7,5 µg/100µl mengalami kematian total berturut-turut setelah 15 dan 19 hari uji tantang, sedangkan ikan kontrol yang tidak divaksin mengalami kematian total setelah 17 hari uji tantang. Jumlah leukosit total ikan yang divaksinasi lebih tinggi dibanding dengan kontrol sampai hari ke-42, setelah itu mengalami penurunan. Indeks fagositosis ikan yang divaksin dengan dosis 12,5 µg/100µl mengalami penurunan setelah hari ke-49 atau 7 hari setelah uji tantang. Kata kunci: Vaksin DNA, Koi herpesvirus (KHV), leukosit, indeks fagositosis, Cyprinus carpio

Effectiveness of B-actin promoter on driving target gene expression in common carp transgenesis Hidayani, Andi Aliah; Carman, Odang; Alimuddin, .
Jurnal Akuakultur Indonesia Vol. 10 No. 1 (2011): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

Promoter in transgene construct plays an important role on regulating of transgene expression level in transgenic fish. In fish transgenesis, researcher convinced that use all-fish gene construct is safety and prospective. This study was performed to compare effectiveness b-actin promoter, - the promoter which has ubiquitous, constitutive, housekeeping characteristics, from common carp (homologous) and from tilapia and medaka b-actin promoters (heterologous) in driving of green fluorescent protein (GFP) expression as a model of target gene on common carp transgenesis. These gene constructs were separately microinjected into cytoplasm of 60 one-cell-stage common carp embryos. The results suggested that 70% survival rate at embryo stage and 45% hatching rate values showed that the microinjection was performed successfully. Percentage of embryos expressing GFP gene were slightly higher when injected using common carp and medaka promoters than those of using tilapia promoter. Percentage of larvae expressing GFP using common carp promoter was similar with medaka promoter. Furthermore, GFP expression using common carp b-actin promoter could be detected at one-week-old larvae, while GFP expressing using medaka b-actin promoter was lasted at 2-day-old larvae. The results demonstrated that homologous promoter more effective in driving of a target gene expression than that of heterologous promoter. Key words: homologous promoter, GFP, transgenesis, common carp ABSTRAK Promoter dalam konstruksi transgen berperan penting dalam pengaturan tingkat ekspresi transgen pada ikan transgenik. Dalam transgenesis ikan, peneliti meyakini bahwa penggunaan konstruksi gen "all-fish" adalah aman dan prospektif. Penelitian ini dilakukan untuk membandingkan efektivitas promoter β-aktin, - promoter yang memiliki ciri ubiquitous, constitutive, dan housekeeping, dari ikan dari ikan mas (homolog) dan ikan nila dan ikan medaka (heterolog) dalam mengendalikan ekspresi gen GFP sebagai model gen pada transgenesis ikan mas. Setiap konstruksi gen tersebut diinjeksikan secara terpisah ke sitoplasma embrio ikan mas fase 1 sel sebanyak 60 embrio. Hasil penelitian dengan kelangsungan hidup embrio 70% dan derajat penetasan 45% menunjukkan bahwa kegiatan mikroinjeksi berhasil dengan baik. Persentase embrio mengekspresikan gen GFP yang diinjeksi konstruksi gen dengan promoter β-aktin ikan mas dan ikan medaka sedikit lebih tinggi dibandingkan dengan yang menggunakan promoter β-aktin ikan nila. Selanjutnya, ekspresi gen GFP yang dikendalikan oleh promoter β-aktin ikan mas dapat dideteksi pada larva berumur 1 minggu, sedangkan ekspresi GFP dengan promoter β-aktin ikan medaka hanya bisa terdeteksi hingga larva berumur 2 hari. Hasil penelitian menunjukkan bahwa promoter homolog adalah lebih efektif dalam mengatur ekspresi gen target dibandingkan dengan promoter heterolog. Kata kunci: promoter homolog, GFP, transgenesis, ikan mas

Production and bioactivity test of recombinant protein common carp growth hormone Utomo, Deny Sapto Chondro; Alimuddin, .; Sudrajat, Agus Oman; Faizal, Irvan
Jurnal Akuakultur Indonesia Vol. 10 No. 1 (2011): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

This study aimed to produce recombinant growth hormone (rGH) protein of common carp (Cyprinus carpio) and evaluate its bioactivity. DNA fragment encoding mature GH protein of common carp (mCcGH) was amplified by polymerase chain reaction (PCR) method and PCR products were then ligated into pCold I to generate pCold I-mCcGH protein expression vector. Escherichia coli BL21 (DE3) harboring pCold I-mCcGH was cultured in the 2xYT medium at 15 °C for 24 hours and protein production was induced by isopropyl-beta-thio galactopyranoside (IPTG). The inclusion bodies containing rGH protein from E. coli transformants were isolated by sonication method and analyzed by SDS-PAGE. The result showed that rGH with molecular weight of about 25 kDa was obtained. Common carp juveniles with average body weight of 5.2±0.4 g were intramuscularly injected once a week for 4 weeks with rGH protein solution from 1 μg bacterial cells per gram fish body weight. The result showed that juveniles fish injected with rGH grew 106.56% higher than control. This result indicated that rGH produced in E. coli BL21 possessed biological activity and it may be useful to improve growth of aquaculture species.Key words: growth hormone, recombinant protein, common carp ABSTRAKPenelitian ini bertujuan menghasilkan protein rekombinan hormon pertumbuhan (growth hormone, GH) dari ikan mas (Cyprinus carpio) dan menguji bioaktivitasnya. Fragmen DNA penyandi protein matang (mature) GH ikan mas (mCcGH) diamplifikasi dengan menggunakan metode PCR dan hasilnya kemudian diligasi ke dalam pCold-I untuk menghasilkan konstruksi vektor ekspresi pCold-I-mCcGH. Plasmid pCold-I-mCcGH ditransformasi ke bakteri Escherichia coli BL21 (DE3), dikultur dalam media 2xYT cair pada suhu 15°C selama 24 jam dan produksi protein diinduksi dengan menggunakan isopropyl-beta-thio galactopyranoside (IPTG). Badan inklusi yang mengandung protein rekombinan GH (rGH) dari bakteri E. coli transforman diisolasi menggunakan metode sonikasi dan dianalisis dengan menggunakan SDS-PAGE. Hasil penelitian menunjukkan bahwa rGH dengan bobot molekul sekitar 25 kDa berhasil diproduksi. Benih ikan mas dengan bobot rata-rata 5,15±0,4 g diinjeksi secara intramuskular satu kali per minggu selama 4 minggu dengan larutan rGH hasil ekstraksi dari 1 µg pelet bakteri/g bobot ikan. Benih yang disuntik dengan rGH tumbuh sekitar 100% lebih cepat bila dibandingkan dengan kontrol yang tidak diinjeksi rGH. Hasil ini mengindikasikan bahwa rGH yang diproduksi dalam bakteri E. coli memiliki bioaktivitas dan dapat bermanfaat untuk memacu pertumbuhan spesies ikan-ikan budidaya.Kata kunci: hormon pertumbuhan, protein rekombinan, ikan mas

Performance of Nile tilapia juvenile fed diet containing different dosages ofrecombinant common carpgrowth hormone Hardiantho, Dian; Alimuddin, .; Prasetiyo, Arief Eko; Yanti, Dwi Hany; Sumantadinata, Komar
Jurnal Akuakultur Indonesia Vol. 11 No. 1 (2012): Jurnal Akuakultur Indonesia
Publisher : Indonesian Society of Scientific Aquaculture (ISSA)

The studywas conductedtodetermine thedose of recombinant common carp growth hormone (rCcGH) supplemented in the commercial diet that generates the best performanceonbodyweight, biomass, andfeed conversion rate of juvenile tilapia.The dose of rCcGH administered was10, 20, and30mg/kg of commercial feed, andnorGH supplementation as control. Eachtreatment was designed in triplicates. Juveniles at average bodylengthof about2cm(average body weight of 0.7 g) were reared in a density of25fish/m2, for three weeks in hapa net at a size of 2×2×1 m3. Fish were fed diet containing rCcGH twicea week with an interval ofthreedays. The juvenile were fed with the rCcGH supplemented diet three times daily at satiation. The results showedthat average body weight andbiomass of fish treated by rCcGH with the doses of 20 and 30 mg/kg feed was higher (p<0.05) than that of 10 mg/kg and control. In addition, feed conversionrate of 20 and 30 mg/kg feed treatments was lower (p<0.05) than that of 10 mg/kg and control. RT-PCR analysisshowedthat expression level of IGF-1 gene in juvenile liver was increased in parallel with the increasing of rCcGH dosages supplemented in diet.This suggestedthatIGF-1 plays an important rolein the induction ofgrowth,andfeed conversionefficiencyof tilapiafed diet containing rCcGH. The bestperformance of juvenile can be obtainedby feeding with diet containingrCcGH20‒30mg/kgof feed. Keywords: rCcGH, IGF-I, recombinant growthhormone,performance, Nile tilapia.

Co-Authors . Irmawati . Kurdianto . Suharsono A. Octavera A. Santika Abdul Halim Talli Abdul Malik Abyadul Fitriah Abyadul Fitriyah Adi Sucipto, Adi Adinda Rizky Tasrianty Afdal Madein Afif Abdurrahman Afpriyaningrum MD Agus Oman Sudrajat Agus Priyadi Ahmad Faisal Ahmad Muzaki Ahmad Ramzi Ghulam Syam Ahmad Zaki Alfiah Mutmainnah Ali Alyaminy, Humaidatul Aminah Zaim Aman Sentosa Panggabean Amin, Muhmmadiyah Andi Aliah Hidayani Andi Haerul Andi Maulana Kamri Andi Parenrengi Andi Parenrengi Andi Parenrengi Andi Tenriulo Andi, K. Petta Lolo, Arjuni Andri Gemaini Angela Mariana Lusiastuti Anggriani, Emi Anhar Anita zafirah ANNA OCTAVERA Anny Hary Ayu Apriwanto, Indra Aras Syazili Ardo, Okilanda Arief Boediono Arief Eko Prasetiyo Arien Heryansyah Asdar Asep Permana Asep Sopian Asri Ainun Amaliah Atradinal Aulia Asman Aulia Saputra Awi Dassa Ayi Santika Azis Ali Baderiah Baderiah Baderiah Bahri, Nur Alim Benyamin Timotiwu, Paul Bohari Yusuf Boyun Handoyo Budimawan Chaeroni, Ahmad Chaerunnisa Chotban, Sippah DARMAWAN SETIA BUDI Darussalam Darwin, Khadijah DEDI JUSADI Delfian Masrura Deny Sapto Chondro Utomo Desy Nurul Astuti Devi Wardan Sari Dian Hardiantho Diki Nurbaldatun Islam Dinar Tri Soelistyowati Djadir Dwi Hany Yanti Eko Pramono Emma Suryati Emma Suryati Enang Harris Enang Harris Endi Permata Eni Kusrini Eni Kusrini Erma Primanita Hayuningtyas F.H. Pasaribu Fachruddin Fachruddin Fajar Anggraeni Fatihah, Anariski Fatimah Akbal Fauzan Wahib Alsani Fauziah Zainuddin Firman Firmansyah Fitriadi Fitriyah Husnul Khotimah Fitry Hasdanita Fredinan Yulianda G. Yoshizaki Gemi Triastutik Gloria Ika Satriani Goro Yoshizaki GORO YOSHIZAKI Gunawan Gunawan Gustikasari Amin H. Arfah Hadi Nasbey Hafidah, Riva Halimah Hamda Hamid Habbe Hamzah Hanipa Hardiantho D Harmayani, Ria Harton Arfah Hary Krettiawan Haryanti Haryanti Hasan Nasrullah Hassan, Muhammad Nasiru Herjayanto, Muh. Hisyam Ihsan Husain , Muh. Saleh Ibnu Izzah Idrus Sahar Muhammad Iis Diatin Ika Kartika Ikhlas Amal Alam, Ade Ikhwanul Arifan ILHAM Ilham Minggi Imron Imron Indah Mastuti Intan Febrianti Irma Andriani IRMA ANDRIYANI Irmawati, Budimawan Irvan Faizal Irvan Faizal Irwan Irzal Effendi Ita Djuwita Jasmadi Jasmadi JATI, M. SASMITO Jean-Christophe Avarre Joko Santoso Jr., Muhammad Zairin Jubaedah Julie Ekasari Jumrah123 K. Chaitanya Deepti K. Sumantadinata Kabangnga, Felix Jansen Kadriani, Nur Annisa Kamaruddin Kartika, Ni Made Andry Kartini, Selvira Febriani Katarina Manik, Tumiar Ketut Mahardika Ketut Sugama Khaerullah khairuzzazi Khayati, Sri Komar Sumantadinata Komar Sumantadinata Kurniawati, Wd. Intan Leman, Swanggi LESTARI DEWI, AYU Lianasari, Ika Yekti Lidyasari Lisa Risdianti Putri Liza Luky Adrianto Luthfy An-nisa Majid M. Zairin Junior M.Y. Jamal Madein, Afdal Maharani, Adinda Mariani, Yuni Marwah, Andi Marwah Mashud MEDIATY Megawati, Novi Mia Setiawati Miftahul Jannah Mira Mawardi MMSI Irfan ,S. Kom Moch. Noer Alim Mochamad Syaifudin, Mochamad Muh Alias L. Rajamuddin Muh. Akmal, Andi Muh. Alias L. Rajamuddin MUHAMMAD AGUS SUPRAYUDI Muhammad Ammar Naufal Muhammad Awal Ramadhan Muhammad Firdaus Muhammad Ikhsan Muhammad Jufri Muhammad Muttaqin Muhammad Sabir Muhammad Safir Muhammad Wawan Muhammad Zairin Jr. Muhammad Zairin Jr. Muhammadiyah Amin Muhram, La Ode Mukni’ah Munawaroh, Zuhrotul MUNTI YUHANA Muslim Muslim Mustakim Mustika Sari, Mustika Musyfikah Ilyas Muzahar N.A. Maswan Nabila Ulfa Nadhirah Nagu Nagu, Nadhirah Nasrullah Nefi Andriana Fajri Ni Made Andry Kartika Nining Asniar Ridzal Noerlitasari Nugraha, Adrianto Fahrizal Nunak Nafiqoh Nur Bambang Priyo Utomo Nur, La Ode Muh.Adam Nur, Rezki Amaliah Nurdin Arsyad Nurfadilla Nurhalija Nurhayati Nurhidayat, Muhammad Nurlaelah Nurlaelah Syarifuddin Nurly Faridah Nurly Faridah Nurul Fadhilah Nurwati Djam’an O. Carman O.Z. Arifin Odang Carman Oman Unju Subandi P. Ratna Kumar Permata Sari, Anggun Petrus Rani Pong-Masak Priatama, Rizki Purwaningsih, Yuli Pustika Ratnawati Putri, Angelina Atika R Rifai R.D. Soejoedono Rahmi, Kurnia Anggraini Ramaita Ratu Siti Aliah Ridwan RIDWAN AFFANDI Risfiyanty, Dwi Kartika Riswandi Rizky Ayu Fatimah Ridwan Rodliyah Rohayu H, Rina Rosidah Rusli RUSTIDJA S. Mariam Salwa Nur Salsabila Samsu Adi Rahman Sani, NurHaslina Sanusi Saputra, Ade Maulana Saputra, Mardepi Sari , Nur Indah Sari Budi Moria Sembiring Sari, Reska Handayani Sarmila Sekar Sulistyaning Hadiwibowo Setiawati M Shabarullah Shalsyabila , Zabina Shandi Irawan Sholeh Ridwan Siddiq, Mustagfirah Siti Aslamyah Siti Fadilah Siti Khodijah Siti Nurul Haliza Sitti Fakhriyyah Sitti Zuhaerah Thalhah Slamet Subyakto Soelistyowati DT Soerja Koesnarpadi Soko Nuswantoro Sri Khayati sri murtini . Sri Nuryati Sri Nuryati Srihartini, Yusi St. Fatmawati. L St. Hidayah Triana St. Mukarramah Sukenda . Sukenda Sukenda Sukmawati Ali Sumardin Raupu Sundari Sutamrin Syafrianto, Syafrianto Syuhada Restu Danupratama T. Takeuchi Tahrim, Tasdin Talib, Dr. Ahmad Tasdin Tahrim Tedy Murtedjo Tri Setyo Guntoro Usman Mulbar Utut Widyastuti Utut Widyastuti Utut Widyastuti UTUT WIDYASTUTI Viona Amelia Candra Wa Ode NurRadhiah Ridjalu Wahidah Sanusi Wartono Hadie Wicaksono, Aryo Wenang WIDANARNI WIDANARNI Y. Hadiroseyani Yulintine Yulintine Yuni Mariani Yuni Puji Hastuti Yuningsih, Andri Yusra, Andi Yusuf, Niken Yulian Zairin Jr M Zakki Zainun

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