Irvan Faizal
Agency for Research Assessment and Application of Technology (BPPT), Serpong.

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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 : Jurnal Akuakultur Indonesia

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Abstract

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   ABSTRAK Penelitian 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
Evaluation of different promoters driving the GFP reporter gene in seaweed Kappaphycus alvarezii Rajamuddin, Muh. Alias L.; Alimuddin, A.; Widyastuti, Utut; Faizal, Irvan
Indonesian Journal of Biotechnology Vol 19, No 2 (2014)
Publisher : Universitas Gadjah Mada

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Abstract

Promoter regulates expression level of foreign gene in transgenic organism. This study was performed to select asuitable promoter as the fi rst step towards production of valuable trait-enhanced seaweed by transgenic technology. Greenfl uorescent protein (GFP) gene was used as a reporter to determine the activity of promoter in seaweed Kappaphycusalvarezii. GFP gene constructs driven by cytomegalovirus (pCMV-GFP), caulifl ower mosaic virus (pCaMV-GFP),medaka β-actin (pmBA-GFP) and Japanese fl ounder keratin (pJfKer-GFP) promoters were introduced by electroporationmethod. Electroporation was performed using a gene pulser (BIORAD) with voltage of 300 V, pulse length of 0.5 ms,pulse numbers of 4, and pulse interval of 0.1 s. Promoter activity was determined by analyzing GFP gene expressionlevel using a fl uorescent microscope. The results showed that CMV regulated highest number of fi lament callus(34.10%±1.49) expressing GFP at medium to strong fl uorescence levels. CaMV promoter had relatively similar activitywith CMV, but lower number of fi lament callus expressing GFP (10.48%±0.25). mBA promoter drove GFP expressionat medium level and similar number of fi lament callus (8.85%±2.31) expressing GFP with CaMV, while JfKer promoterhad lowest activity by means in number of fi lament callus expressing GFP (4.79%±0.26) and GFP expression level. PCRanalysis for transgenic confi rmation showed a DNA band of PCR product from pCMV-GFP and pCaMV-GFP expressingfi lament callus in the same size (about 0.6 kb) with positive control of plasmid. Thus, CMV and CaMV promoters wasan appropriate promoter and foreign gene could be transferred to fi lament callus by electroporation method. Combiningthis achievement with developing a culture method of fi lament callus to be thallus, stable transgenic breeding in K.alvarezii can be feasible.
IDENTIFICATION OF MAJALAYA COMMON CARP STRAINS RESISTANT TO KHV INFECTION USING CYCA-DAB1*05 ALLELE AS THE MARKER Alimuddin, Alimuddin; Mubinun, Mubinun; Santika, Ayi; Carman, Odang; Faizal, Irvan; Sumantadinata, Komar
Indonesian Aquaculture Journal Vol 6, No 2 (2011): (December 2011)
Publisher : Pusat Penelitian dan Pengembangan Perikanan, Badan Penelitian dan Pengembangan Kelautan da

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Abstract

The Cyca-DAB1*05 allele of major histocompatibility complex class II genes is recently suggested to have a link with the European common carp strain resistant to koi herpesvirus (KHV). In this study, a set of specific primers for Cyca-DAB1*05 was designed and applied as a marker to identify broodstocks of majalaya common carp strain subsequently used as a candidate resistant to KHV infection. From a total of 23 broodstock subjected to PCR analysis, two female and male fish, both having (P) and no Cyca-DAB1*05 (N), were selected and then diallelly mated. Disease resistance of progenies from 10 crosses was determined by a survival analysis in pond rearing and a laboratory challenge-test using cohabitation method. The results have revealed that the average survivals of PxP progenies for pond rearing and KHV challenge test were 86% and 100% higher (P < 0.05) respectively compared to that of NxN fish. Survival rate of PxN/NxP progenies was significantly lower (P < 0.05) than that of PxP fish. Furthermore, PCR analysis showed that almost 91% progenies of PxP crosses seemed to have a KHV resistant gene marker. Thus, this study suggests that the marker is associated with the KHV resistance in majalaya common carp strain, and farming of PxP progenies can be useful to increase common carp production.
PRODUKSI PROTEIN REKOMBINAN HORMON PERTUMBUHAN IKAN KERAPU Faizal, Irvan; Aliah, Ratu Siti; Amarullah, Muhammad Husni; Megawati, Novi; Sutanti, Sutanti; Alimuddin, Alimuddin
Jurnal Riset Akuakultur Vol 7, No 2 (2012): (Agustus 2012)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

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Abstract

Salah satu spesies ikan yang menjadi target produksi perikanan budidaya nasional adalah ikan kerapu tikus (Cromileptes altivelis). Ikan kerapu tikus merupakan ikan laut budidaya komoditas ekspor, namun laju pertumbuhannya sangat lambat. Oleh karena itu, perlu dilakukan usaha budidaya yang mampu menaikkan laju pertumbuhan ikan kerapu tikus. Pendekatan nutrisi melalui penggunaan hormon pertumbuhan (Growth Hormone, GH) pada usaha budidaya diyakini mampu meningkatkan kecepatan tumbuh ikan budidaya. Pada penelitian ini, melalui teknologi DNA rekombinan telah dilakukan isolasi dan identifikasi gen GH yang selanjutnya dilakukan produksi protein rekombinan GH (recombinant Growth Hormone, rGH) dengan memanfaatkan penggunaan bakteri E. coli. Dalam pelaksanaan penelitian dilakukan konstruksi rGH yang menghasilkan bakteri E. coli BL21 (DE3) yang mampu memproduksi protein rGH. Produksi rGH dilakukan pada skala bioreaktor. Proses isolasi produk rGH-nya dalam bentuk pellet inclusion bodies yang selanjutnya dicampur dengan pelet pakan komersil hingga konsentrasi akhir protein dalam pakan mencapai 1 ng/6 mg pakan, di mana setelah dikering-anginkan, pelet pakan protein rekombinan GH dapat diaplikasikan untuk budidaya ikan kerapu tikus dan ikan budidaya lainnya.
Pemberian hormon pertumbuhan rekombinan secara “putus dan sambung” pada tiga kelompok ukuran benih ikan kerapu bebek, Cromileptes altivelis (Valenciennes 1828) [“Stop and go” treatment of recombinant growth hormone to different sizes of humpback grouper juveniles, Cromileptes altivelis (Valenciennes 1828)] Antoro, Suci; Alimuddin, nFN; Suprayudi, Muhammad Agus; Faizal, Irvan; Junior, Muhammad Zairin
Jurnal Iktiologi Indonesia Vol 15, No 1 (2015): Februari 2015
Publisher : Masyarakat Iktiologi Indonesia (MII )

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Abstract

The objective of this study was to compare the growth response of 3 different sizes of humpback grouper juveniles, which were come from the same brood stock group and spawning season after “stop and go” experiment of recombinant Epinephelus lanceolatus growth hormone (rElGH); that are with, without and retreated with rElGH for 42 days respectively. Each size group was divided into two treatment groups. The first group was treated with 50 mg crude rElGH kg-1 in commercial diet (pC) and a second group as a control. Weight gain of pC compares to control for small size group, medium size group and large size groups of juveniles subsequently for first stage were 85.89%, 39.66% and 16.34%; second stage were -34.57%, -14.76%, and -5.27%; and third stage were 56.16%, 50.24% and 59.14%. Specific growth rate differences of small, medium and large size of pC compared to control in first stage were 41.6%, 19.06% and 7.52%; second stage were -44.81%, -27.23% and -14.66%; and third stage were 55.9%, 40.62% and 48.42%. No significant difference of condition factor among all sizes of pC and control fish. Protein content and retention, and liver glycogen content from pooled sample of all size fish groups pC treatment in the second stage were decreasing compared to the first stage, respectively, 11.49%, 35.14% and 84.73%. It can be concluded that rElGH treatment improved growth performance of all size fish groups, however small juveniles have highest growth response compared to medium and large juvenile groups. The ceasing of rElGH treatment on second experiment stage is most likely causing the loss of accelerating growth factor then decreasing growth performance, protein content and retention, and liver glycogen content. AbstrakPenelitian bertujuan membandingkan respons pertumbuhan tiga kelompok ukuran benih ikan kerapu bebek dari kelompok induk dan periode pemijahan yang sama terhadap hormon pertumbuhan rekombinan ikan kerapu kertang Epine-phelus lanceolatus (rElGH); melalui eksperimen “putus dan sambung” yaitu dengan, tanpa, dan perlakuan kembali rElGH masing-masing selama 42 hari. Setiap kelompok ukuran dibagi menjadi dua kelompok perlakuan, kelompok pertama diberi perlakuan rElGH dengan dosis 50 mg rElGH-HP55 kg-1 pakan (pC) sedangkan kelompok kedua sebagai kontrol. Pertambahan bobot badan kelompok pC dibandingkan dengan kontrol pada benih berukuran kecil, sedang dan besar berturut-turut pada eksperimen tahap pertama 85,89%, 39,66% dan 16,34%; tahap kedua -34,57%, -14,76%, dan -5,27%, dan tahap ketiga 56,16%, 50,24% dan 59,14%. Perbedaan laju pertumbuhan spesifik benih berukuran kecil, se-dang dan besar perlakuan pC terhadap kontrol pada eksperimen tahap pertama 41,6%, 19,06% dan 7,52%; tahap kedua -44,81%, -27,23% dan -14,66%; dan tahap ketiga 55,9%, 40,62% dan 48,42%. Faktor kondisi pC dan kontrol pada se-mua kelompok ukuran tidak berbeda nyata. Kandungan dan retensi protein, dan kandungan glikogen hati gabungan sampel dari semua kelompok ukuran ikan perlakuan pC pada eksperimen tahap kedua menurun dibandingkan eksperi-men tahap pertama, masing-masing sebesar 11,49%, 35,14% dan 84,73%. Dapat disimpulkan pemberian rElGH mema-cu pertumbuhan semua kelompok ukuran benih ikan, namun benih berukuran kecil mempunyai respons pertumbuhan lebih tinggi daripada kelompok benih berukuran sedang dan besar. Penghentian pemberian rElGH menyebabkan ber-hentinya faktor pemacu pertumbuhan, sehingga performa pertumbuhan, kandungan dan retensi protein, dan kandungan glikogen hati menurun.
Analysis of flowering gene in palm oil (Elaeis guineensis) FAIZAL, IRVAN; EMDI, AXEL
Asian Journal of Agriculture Vol 1 No 02 (2017)
Publisher : Society for Indonesian Biodiversity & Universitas Mulawarman, Samarinda, Indonesia

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Abstract

Faizal I, Emdi A. 2017. Analysis of flowering gene in palm oil (Elaeis guineensis). Asian J Agric 1: 53-58. Palm oil has always been an important commodity in Indonesia. The most common species is palm oil, Elaeis guineensis. Palm oil is a monoecious plant with a tendency to be a temporal dioecious. Female flower will be the one that produces palm oil fruit, that later is treated with palm oil while male flower only takes part in the fertilization process. In order to know the ratio between female and male flower tree in a plantation, this study was performed to detect a distinction between female and male flowering gene sequences from DNA sample of E. guineensis. Based on previous study which managed to characterize MADS-box gene of palm oil, a primer was designed and named GmG (Globosa-male-Gaps). The result shows that the primer has the ability to differentiate DNA sequence female and male flower of E.guineensis, Palm oil. However, further studies with full sequence and more samples are needed to find distinctive results between female and male flower sequences as the GmG primer could be used to design a specific marker or primer to detect the presence of female or male flower within a tree.