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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 : Jurnal Akuakultur Indonesia

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Abstract

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
Pemanfaatan tepung testis sapi sebagai hormon alami pada penjantanan ikan cupang, Betta splendens Regan, 1910 [Cow’s testicles flour as the natural hormone masculinization of Siamese fighting fish, Betta splendens Regan, 1910] Hidayani, Andi Aliah; Fujaya, Yushinta; Trijuno, Dody Dharmawan; Aslamyah, Siti
Jurnal Iktiologi Indonesia Vol 16, No 1 (2016): Februari 2016
Publisher : Masyarakat Iktiologi Indonesia (MII )

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Abstract

Siamese fighting fish, Betta splendens male is a lovely color ornamental fish with unique shape fins that make it highly demand by the ornamental fish lovers. This study aims to perform sex reversal with masculinization fish production. The study was carried out in two stages i.e.: stage 1 by soaking the 4 days old fish larvae into a solution of cow testicles flour with different doses, stage 2 with different soaking time. Testicular dose tested consists of five levels i.e.: 0 mg L', 20 mg L-1, 40 mg L-1, 60 mg L-1, and 80 mg L-1. Time immersions tested were: 0 hours, 24 hours, 36 hours, 48 hours and 60 hours. The measured parameter was the percentage of male fish produced. The results showed the highest per-centtage of male fish obtained at a dose of 60 mg L-1 and a 24-hour soaking time with a percentage value respectively 88.5% and 87.5%. The study provided information that masculinization technology in a solution of cow testicles applicable for fish larvae. This technology is easy to do so that farmers can use cow's testicles flour for masculinization for their fish production. AbstrakIkan Cupang, Betta splendens jantan merupakan ikan hias yang memiliki keindahan warna tubuh serta keunikan bentuk sirip sehingga sangat diminati oleh pecinta ikan hias. Penelitian ini bertujuan melakukan pembalikan kelamin dengan menjantankan ikan cupang yang diproduksi. Penelitian dilakukan dalam dua tahap yaitu: tahap pertama dengan meren-dam larva ikan cupang berumur empat hari ke dalam larutan tepung testis sapi dengan dosis berbeda, dan tahap ke dua dengan lama perendaman berbeda. Dosis testis yang diuji terdiri atas lima tingkatan yaitu 0 mg L-1, 20 mg L-1, 40 mgL-1 60 mg L-1, dan 80 mg L-1. Lama perendaman yang diuji adalah: 0 jam, 24 jam, 36 jam, 48 jam, dan 60 jam. Parameter yang diukur adalah persentase ikan jantan yang dihasilkan. Hasil penelitian menunjukkan bahwa persentase ikan berke-lamin jantan tertinggi diperoleh pada dosis 60 mg L-1 dan lama waktu perendaman 24 jam dengan nilai persentasi ber-turut-turut 88,5% dan 87,5%. Hasil penelitian ini memberikan informasi bahwa teknologi penjantanan melalui perendaman dalam larutan testis sapi dapat dilakukan pada larva ikan cupang. Teknologi ini mudah dilakukan sehingga pembudidaya dapat menggunakan tepung testis sapi untuk menjantankan ikan cupang produksinya.
Kloning promoter P-actin ikan mas, Cyprinus carpio Lin. 1758 dan analisis fungsionalnya menggunakan gen target protein pendaran hijau (GFP) [P-actin promoter cloning of common carp, Cyprinus carpio Lin. 1758 and its functional analysis using targeted Green Fluorescent Protein (GFP) gene] Hidayani, Andi Aliah; Carman, Odang; Alimuddin, nFN
Jurnal Iktiologi Indonesia Vol 13, No 2 (2013): Desember 2013
Publisher : Masyarakat Iktiologi Indonesia (MII )

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Abstract

Promoter in the expression vector plays an important role on regulating of gene expression in transgenic fish. In fish transgenesis, researcher convinced that the use of all-fish expression vector is safety and prospective. The study was performed to isolate P-actin promoter, the promoter which has ubiquitous, constitutive, housekeeping characteristics, of common carp as a first step to construct all-common carp expression vector. Common carp P-actin promoter (ccBA) was isolated using PCR method with FBP1, RBP1, and RBP2 primers. Sequencing was performed using ABI PRISM 3100 machine, and analysis of sequences was conducted using GENETYX version 7 software. The results of sequence analysis showed that the length of DNA fragment obtained was approximately 1.5 kb. Results of homology with P-actin promoter sequence of a gene bank database (Accession No.: M24113) was 97.5%. The evolutionary conserved of transcription factor for P-actin promoter including CCAT, CArG, and TATA boxes were found in the sequence. Ubiquitous and higher expression of green fluorescent protein driven by ccBA promoter in muscle of common carp larvae was detected. It is most likely that the isolated sequence is a common carp P-actin promoter. AbstrakPromoter dalam vektor ekspresi berperan penting dalam mengatur ekspresi gen pada ikan transgenik. Dalam transgenesis ikan, peneliti yakin bahwa penggunaan vektor ekspresi semua ikan aman dan prospektif. Penelitian ini dilakukan untuk mengisolasi promoter P-aktin, promoter yang memiliki karakteristik ubiquitous, constitutive, house keeping, dari ikan mas sebagai langkah awal untuk mengkonstruksi vektor ekspresi semua ikan mas. Promoter P-aktin ikan mas (ccBA) diisolasi menggunakan metode PCR dengan primer FBP1, RBP1, dan RBP2. Sequensing dilakukan dengan menggunakan mesin ABI PRISM 3100, dan analisis sekuen dilakukan dengan menggunakan perangkat lunak GENE-TYX versi 7. Hasil analisis sekuen menunjukkan bahwa panjang fragmen DNA yang diperoleh adalah sekitar 1,5 kb. Hasil homologi dengan sekuen promoter P-aktin dari pangkalan data bank gen (No. Aksesi: M24113) adalah sebesar 97,5%. Faktor transkripsi yang tetap secara evolusioner untuk promoter P-aktin promoter termasuk CCAT, CArG, dan boks TATA ditemukan dalam sekuen. Ubiquitous dan ekspresi tertinggi protein pendaran hijau (GFP) dikendalikan oleh promoter ccBA dalam otot larva ikan mas yang dideteksi. Dengan demikian, kemungkinan besar bahwa sekuen yang terisolasi adalah promoter P-aktin ikan mas.
Masculinization of guppy fish (Poecilia reticulata Peters, 1859) with extract of sea cucumber (Holothuria scabra) Saputra, Anugerah; Wulandari, Astri; Ernawati, .; Yusuf, Muhammad Amri; Eriswandy, Irvan; Hidayani, Andi Aliah
Jurnal Iktiologi Indonesia Vol 18, No 2 (2018): June 2018
Publisher : Masyarakat Iktiologi Indonesia (MII )

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Abstract

Guppy fish Poecilia reticulata is one of the most popular freshwater ornamental fish species because it is easy to main-tain, and has beautiful variations of color, especially males. One method to produce male guppy is masculinization me-thod. This study aimed to determine the dose of hormone treatment from visceral organ of sea cucumber to the process masculinization of fish guppy. This study used a completely randomized design with the immersion treatment of parent guppy which has been bred into the extract solution of sea cucumber innards for 24 hours with different doses i.e. 0 mg L-1, 10 mg L-1, 20 mg L-1, 30 mg L-1 and 22 mg L-1 (metiltestosterone as positive control). The parameter measured was the percentage of male fish produced. The results showed that although the effect of the treatment was not significantly different, but it can be said to be significantly different to the treatment of 0 mg L-1 (control) to obtain the best result for masculinization of guppy at 30 mg L-1 immersion and the best survival rate on treatment of 0 mg L-1 (control). The re-sults of this study suggest that the masculinization technology on the parent stock can be done using extracts of sea cu-cumber innards which has been considered as waste.AbstrakIkan gapi Poecilia reticulata merupakan salah satu jenis ikan hias air tawar yang banyak digemari karena mudah dipe-lihara, dan memiliki variasi wama yang indah, terutama jantannya. Salah satu metode untuk menghasilkan ikan gapi jantan adalah dengan metode pembalikan kelamin melalui produksi penjantanan ikan gapi. Penelitian ini bertujuan un-tuk menentukan dosis hormon perlakuan ekstrak jeroan teripang pasir terhadap proses penjantanan ikan gapi. Penelitian  ini menggunakan rancangan acak lengkap dengan perlakuan perendaman induk ikan gapi yang telah dipijahkan ke dalam larutan ekstrak jeroan teripang pasir selama 24 jam dengan dosis berbeda, yaitu 0 mg L-1, 10 mg L-1, 20 mg L-1, 30 mg L-1 dan 22 mg L-1 (metiltestosteron sebagai kontrol positif). Parameter yang diukur adalah persentase ikan jantan yang dihasilkan. Hasil penelitian menunjukkan bahwa walaupun pengaruh antarperlakuan yang diberikan tidak berbeda nyata, tetapi dapat dikatakan berbeda nyata terhadap perlakuan 0 mg L-1 (kontrol) sehingga diperoleh hasil terbaik untuk penjantanan ikan gapi pada dosis perendaman 30 mg L-1  dan sintasan terbaik pada perlakuan 0 mg L-1 (kontrol). Hasil penelitian ini memberikan informasi bahwa teknologi penjantanan pada induk ikan gapi dapat dilakukan dengan me-manfaatkan ekstrak jeroan teripang pasir yang selama ini dianggap sebagai limbah.
The Morphometric Character and Mitochondrial 16S rRNA Sequence of Portunus pelagicus Fujaya, Yushinta; Hidayani, Andi Aliah; Asphama, Andi Ivo; Trijuno, Dody Dharmawan; Tenriulo, Andi; Parenrengi, Andi
Aquacultura Indonesiana Vol 16, No 1 (2015): Volume 16 Issue 1 Year 2015
Publisher : Indonesian Aquaculture Society (MAI)

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Abstract

Blue swimming crab (Portunus pelagicus) colors have variations from different habitats and they have different vitality. The purpose of this study was to analyze the diversity of the P. pelagicus using morphometric and 16SrRNA mitochondrial sequence.  There were three groups of blue swimming crab characterized by differences in color and density patterns of spots.  The result of Canonical discriminant analysis showed that there were morphometric differences among the three groups.  The main distinguishing morphometric characteristics are the ratio of carapace width and length as well as the ratio of the length and width of cheliped merus.  The third group of crab is also has similarity about 98-99% by the Portunus pelagicus 16S rRNA sequences available in gene banks.  These studies had concluded that differences in color and pattern of the spot density have also morphometric and genetic differences. At least there were two groups based on genetic variation of Portunus pelagicus from Barru Regency waters which may have a variety of different properties.  Further research is needed to study the adaptation abilities of the two groups of crab on the condition of aquaculture as a potential domestic animal.Keywords: Crab; Portunus pelagicus; Morphometric; 16S rRNA.