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"ABSTRAKAir rebusan jahe telah berhasil digunakan sebagai reduktor untuk biosintesis nanopartikel perak. Biosintesis dilakukan dengan mencampurkan air rebusan rimpang jahe dan larutan AgNO3 yang kemudian diinkubasi selama 24 jam. Karakterisasi larutan hasil reaksi dilakukan dengan fotografi warna larutan dan spektroskopi UV-Vis. Variabel preparasi yang diteliti adalah varietas jahe, fase pertumbuhan, preparasi simplisia, dan perlakuan mekanik. Metode ini digunakan untuk meneliti pengaruh variabel preparasi terhadap nanopartikel perak yang dihasilkan. Varietas jahe yang diteliti adalah jahe gajah, jahe merah, dan jahe emprit. Fase pertumbuhan rimpang yang diteliti adalah bagian anakan dan indukan rimpang. Pengaruh metode preparasi simplisia yang yang diteliti adalah efek bentuk rimpang berupa bubuk dan irisan. Perlakuan mekanik pada larutan saat biosintesis dibedakan antara tanpa pengadukan dan dengan pengadukan. Kemudian diteliti juga pengaruh asam askorbat pada reaksi pembentukan nanopartikel perak. Hasil fotografi menunjukkan bahwa larutan berubah warna dari bening ke kuning kecokelatan yang sesuai dengan warna larutan nanopartikel perak. Spektrum UV-Vis larutan mempunyai nilai absorbansi di panjang gelombang sekitar 420 nm yang bertepatan dengan nilai panjang gelombang absorbansi nanopartikel perak, dengan demikian rimpang jahe dapat digunakan sebagai reduktor biosintesis nanopartikel perak. Di antara varietas jahe, jahe gajah menghasilkan nanopartikel perak yang paling baik, yaitu mempunyai nilai absorbansi 2,2 ± 0,4. Berdasarkan hasil karakterisasi, variabel preparasi yang baik di eksperimen ini adalah penggunaan anakan rimpang sebagai bahan dasar simplisia. Metode preparasi dengan irisan rimpang lebih baik daripada dengan menggiling rimpang, dan perlakuan tanpa pengadukan larutan selama proses reaksi menghasilkan kualitas nanopartikel perak yang lebih baik. Penambahan asam askorbat saat reaksi dapat memperbanyak nanopartikel perak yang dihasilkan.

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ABSTRACTInfution water of ginger has been successfully used as a reductant for biosynthesis of silver nanoparticles. Biosynthesis made ​​by mixing infution water of ginger rhizome and AgNO3 solution then incubated for 24 hours. Characterization of the resulting solution is performed in the color photography solution and UV-Vis spectroscopy. Preparation variables studied were varieties of ginger: gajah ginger, red ginger, and emprit ginger, growth phase: tillers and main rhizomes, form of botanicals material rhizomes: powder and slices, mechanical treatment: mixture solution without stirring and with stirring, and addition of ascorbic acid. The results showed that the photographic color solution changes from clear to yellow brownish that matches the color solution of the silver nanoparticles. UV-Vis spectrum of the solution has a absorbance value at about 420 nm wavelength which coincides with the wavelength of the absorbance value of silver nanoparticles, thus the ginger rhizome can be used as a reductant for biosynthesis of silver nanoparticles. Among the varieties of ginger, gajah ginger produce silver nanoparticles which were the best, which has an absorbance value of 2.2 ± 0.4. Based on characterization results, good preparation variable in this experiment is the use of the tiller rhizomes as the botanicals material rhizomes. Preparation botanicals material rhizomes by slicing ​​better than by grinding, and treatment without stirring the solution during the reaction produce quality of silver nanoparticles better. The addition of ascorbic acid can increase the silver nanoparticles product."

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Chikungunya merupakan penyakit menular yang bersifat re-emerging atau penyakit lama yang dapat tersebar kembali. Penyakit yang disebabkan virus chikungunya ini memiliki manifestasi klinis non-spesifik sehingga dibutuhkan metode diagnosis yang cepat dan akurat. Protein E2 yang dikode oleh gen E2 pada virus chikungunya berperan penting sebagai pengikatan reseptor sehingga berpotensi untuk digunakan dalam proses diagnosis penyakit chikungunya. Penelitian ini bertujuan menghasilkan protein rekombinan E2 sebagai bahan dasar produksi antibodi monoklonal yang akan digunakan dalam pengembangan perangkat diagnostik penyakit chikungunya. Metode penelitian yang dilakukan mencakup pembentukan plasmid rekombinan pPICZaA-E2, transformasi plasmid rekombinan pada sel inang Escherichia coli, analisis koloni transforman, transformasi plasmid rekombinan pada sel inang ekspresi Pichia pastoris X-33, analisis fenotipe, hingga ekspresi protein rekombinan E2. Hasil penelitian menunjukkan 281 koloni E. coli transforman pPICZaA-E2 dapat tumbuh pada medium yang mengandung antibiotik zeocin. Hasil analisis PCR koloni transforman menunjukkan gen E2 dengan ukuran 1.260 bp berhasil ditransformasikan ke sel inang E. coli menggunakan vektor pICZaA dengan ukuran 3.569 bp. Hasil analisis PCR genom berhasil mengamplifikasi gen AOX1 berukuran 2,2 kb dan 1,8 kb yang menunjukkan plasmid rekombinan berhasil terintegrasi pada genom P. pastoris dan menghasilkan fenotipe Mut⁺. Pita protein berukuran 40 kDa pada hasil SDS-PAGE menunjukkan protein E2 berhasil terekspresi.

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Chikungunya is known as an infectious, re-emerging disease. Because of the non-specific clinical manifestation, chikungunya needs a rapid and accurate diagnostic method for its detection. Envelope 2 (E2) protein coded by E2 gene in the genome of the virus has an important role as an attachment receptor to a cell that made it potential to be used for diagnosis. This research is aimed to obtain E2 recombinant protein as a basic material for monoclonal antibody production in chikungunya rapid diagnostic test kit development. Chikungunya E2 gene is amplified and ligated with pPICZaA vector to make recombinant DNA clones from E. coli. The clones then isolated and used for protein expression in P. pastoris. The result shows 281 transformants of E. coli colonies can grow on a selection medium that contains zeocin. Analysis of direct colony PCR show E2 gene was transformed and cloned using pPICZaA vector. Analysis of genomic PCR shows 2,2 kb and 1,8 kb bands formed that indicate AOX1 gene was amplified and the integration of recombinant plasmid pPICZaA to P. pastoris genome was successful. Visualization of protein electrophoresis shows protein band was formed at the size of40 kDa that indicate the protein expression was successful.

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"[Penelitian mengenai karakterisasi morofologi bunga dan pollinaria 14 spesies Hoya atau bunga lilin telah dilakukan dari Februari--Mei 2015. Penelitian bertujuan untuk mendeskripsikan dan membandingkan 14 spesies Hoya koleksi Kebun Raya Bogor (KRB) berdasarkan morfologi bunga dan pollinaria. Sampel yang digunakan merupakan 14 spesimen awetan basah bunga koleksi KRB. Data yang diambil berupa data kualitatif, kuantitatif, dan visual melalui metode pengamatan langsung. Hasil penelitian menunjukkan variasi bentuk dan ukuran bunga serta pollinaria pada 14 spesies Hoya koleksi KRB. Hasil juga menunjukkan adanya karakter pembeda antarspesies yaitu bentuk calyx, keberadaan trikom pada bagian tepi calyx, bentuk corolla, bentuk corona, pelengkap anther, bentuk pollinia, bentuk corpusculum, ada atau tidaknya caudicle, bentuk caudicle, ada atau tidaknya pellucid edge, dan bentuk translator.;Research on morphological flowers and pollinaria characterization from 14 species Hoya or wax flowers was conducted from Februari--Mei 2015. The aims of this research was to describe and compare 14 species of Hoya collections from Bogor Botanical Garden (BBG) based on flower and pollinaria morphology. The research was carried out using preserved 14 specimen collections from BBG. The qualitative, quantitative, and visual data were collected by direct observation method. The result showed that the shape and size characters from flowers and pollinaria of 14 Hoya species from BBG were varied. Beside variation of Hoya flowers and pollinaria, there were also some distinguish characters of calyx shape, presence or absence of trichome in calyx, corolla shape, corona shape, anther appendages, pollinia shape, corpusculum shape, presence or absence of caudicle, caudicle shape, presence or absence of pellucid edge , and translator shape.;Research on morphological flowers and pollinaria characterization from 14 species Hoya or wax flowers was conducted from Februari--Mei 2015. The aims of this research was to describe and compare 14 species of Hoya collections from Bogor Botanical Garden (BBG) based on flower and pollinaria morphology. The research was carried out using preserved 14 specimen collections from BBG. The qualitative, quantitative, and visual data were collected by direct observation method. The result showed that the shape and size characters from flowers and pollinaria of 14 Hoya species from BBG were varied. Beside variation of Hoya flowers and pollinaria, there were also some distinguish characters of calyx shape, presence or absence of trichome in calyx, corolla shape, corona shape, anther appendages, pollinia shape, corpusculum shape, presence or absence of caudicle, caudicle shape, presence or absence of pellucid edge , and translator shape., Research on morphological flowers and pollinaria characterization from 14 species Hoya or wax flowers was conducted from Februari--Mei 2015. The aims of this research was to describe and compare 14 species of Hoya collections from Bogor Botanical Garden (BBG) based on flower and pollinaria morphology. The research was carried out using preserved 14 specimen collections from BBG. The qualitative, quantitative, and visual data were collected by direct observation method. The result showed that the shape and size characters from flowers and pollinaria of 14 Hoya species from BBG were varied. Beside variation of Hoya flowers and pollinaria, there were also some distinguish characters of calyx shape, presence or absence of trichome in calyx, corolla shape, corona shape, anther appendages, pollinia shape, corpusculum shape, presence or absence of caudicle, caudicle shape, presence or absence of pellucid edge , and translator shape.]"

"Telah dilakukan penelitian yang bertujuan memperoleh identitas dua isolat bakteri termofilik dari geiser. Isolat LC2-23 diperoleh dari serasah pada geiser di Cisolok, Jawa Barat, Indonesia, dan isolat RKB-2 diperoleh dari serasah pada geiser di Onikobe, Miyagi, Jepang.!!Identifikasi dilakukan berdasarkan gabungan data fenotipik dan genotipik. Berdasarkan karakterisasi fenotipik, isolat LC2-23 memiliki sel berbentuk batang; menghasilkan endospora; motil; gram positif; bersifat aerob dan fakultatif aerob; mampu tumbuh pada suhu 60 oC, sedangkan suhu optimum pertumbuhan 50 oC. Berdasarkan karakterisasi genotipik, data full sequence gen 16S rRNA isolat LC2-23 memiliki homologi 99,1% terhadap Brevibacillus agri. Berdasarkan data fenotipik dan genotipik, isolat LC2-23 diidentifikasi sebagai Brevibacillus agri (Family Paenibacillaceae, Order Bacillales, Class Bacilli, Phylum Firmicutes). Berdasarkan karakterisasi fenotipik, isolat RKB-2 membentuk miselium vegetatif dan aerial yang bercabang; menghasilkan spora aerial; gram positif; bersifat aerob; mampu tumbuh pada suhu 60 oC, sedangkan suhu optimum pertumbuhan 50 oC. Berdasarkan karakterisasi genotipik, data full sequence gen 16S rRNA isolat RKB-2 memiliki homologi yang rendah, yaitu 98,4% terhadap spesies terdekatnya, Thermosporothrix hazakensis (Family Thermosporotrichaceae, Order Ktedonobacteriales, Class Ktedonobacteria, Phylum Chloroflexi). Hasil analisis filogenetik menunjukkan posisi isolat RKB-2 terpisah dari T. hazakensis. Data kemotaksonomi (komposisi asam lemak) dan hasil analisis proteomik menggunakan MALDI-TOF MS mendukung perbedaan antara isolat RKB-2 dan T. hazakensis. Berdasarkan perbedaan tersebut isolat RKB-2 diidentifikasi sebagai spesies baru dari Thermosporothrix. Untuk pengajuan nama spesies baru diperlukan data hibridisasi DNA-DNA antara isolat RKB-2 dengan T. hazakensis.

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This research was aimed to identify two bacterial isolates obtained from geysers. Strain LC2-23 was isolated from litters on a geyser in Cisolok, West Java, Indonesia, and isolate RKB-2 was obtained from litters on a geyser in Onikobe, Miyagi Prefecture, Japan. Identification of bacteria was based on integrated data of phenotypic and genotypic characterizations. Based on phenotypic characterizations of isolate LC2-23: it has a rod (bacilli)-shaped cells, forms endospores; gram positive; motile; aerobic, and able to grow up to a temperature of 60 oC. Based on genotypic characterizations of isolate LC2-23: the full sequence of genes 16S rRNA shows 99.1% sequence homology to Brevibacillus agri. Based on phenotypic and genotypic data, isolate LC2-23 can be identified as Brevibacillus agri (Family Paenibacillaceae, Order Bacillales, Class Bacilli, Phylum Firmicutes). Based on phenotypic characterizations of isolate RKB-2: vegetative and branching aerial mycelia forms, gram positive, aerobic, and able to grow up to a temperature of 60 oC. Based on genotypic characterizations of isolate RKB-2: the full sequence of 16S rRNA gene of isolate RKB-2 showed low homology (98.4%) to Thermosporothrix hazakensis (Family Thermosporotrichaceae, Order Ktedonobacteriales, Class Ktedonobacteria, Phylum Chloroflexi). Phylogenetic analysis showed the isolate RKB-2 was distinct from cluster of Thermosporothrix hazakensis and Ktedonobacteria bacterium. The genotypic and phylogenetic data, plus chemotaxonomic and proteomic analysis using MALDI-TOF MS, suggest that isolate RKB-2 represent novel species of the genus Thermosporothrix. The DNA-DNA hibridization data is needed for proposal of new species."

"[Pleurotus ostreatus atau jamur tiram merupakan salah satu cendawan yangdikonsumsi di Indonesia karena tingginya cita rasa dan nilai nutrisi, serta dapatdibudidaya dengan mudah dan murah menggunakan serbuk kayu sebagai mediatumbuh. Penelitian bertujuan untuk meneliti pengaruh penambahan A. fumigatusdalam proses pengomposan substrat serbuk kayu untuk media tumbuh P. ostreatusterhadap kualitas kompos dan produksi tubuh buah. Hasil penelitianmenunjukkan bahwa suhu, konsentrasi glukosa, xilosa, N-asetilglukosaminmengalami kenaikan, sedangkan nilai pH, kadar selulosa, dan hemiselulosamengalami penurunan, pada hari ke-0 hingga hari ke-7. Berdasarkan databiokimia tersebut, penambahan A. fumigatus pada saat proses pengomposanmeningkatkan kualitas kompos substrat serbuk kayu. Rata-rata kecepatanpertumbuhan miselia per hari pada kelompok perlakuan (1,10 cm) lebih cepatdibandingkan dengan kontrol (1,07 cm) selama 24 hari pengamatan. Analisis ujistatistik ANAVA terhadap berat segar tubuh buah P. ostreatus menunjukkan hasilyang berbeda nyata secara signifikan (α=0,05) tetapi diameter tudung tidakberbeda. Penambahan A. fumigatus dalam proses pengomposan serbuk kayumeningkatkan tubuh buah hasil panen P. ostreatus.;Oyster mushroom Pleurotus ostreatus is the most popular edible mushroom in Indonesia because it is delicious and nutritious and can be cultivated easily and inexpensively using sawdust as the substrate The consumption of oyster mushroom is largely because of its taste and nutritional properties The aims of this research were to reveal the effect of A fumigatus addition in composting process using sawdust substrate for P ostreatus growth medium toward compost quality and yield productivity The experiment revealed that the temperature concentration of glucose xylose and N acetylglucosamine increased and the pH value percentage of cellulose and hemicellulose decreased during 7 days of composting process Based on that biochemical parameter addition of A fumigatus during composting process increased sawdust compost quality Mycelia growth rate per day in treatment group 1 10 cm was faster than control group 1 70 cm during 24 days of observation Statistical test analysis using ANAVA for the fresh weight of P ostreatus indicated that the result was significantly different 0 05 but had no significantly different in diameter of the cap Addition of A fumigatus in sawdust composting process increased yield productivity of P ostreatus , Oyster mushroom Pleurotus ostreatus is the most popular edible mushroom in Indonesia because it is delicious and nutritious and can be cultivated easily and inexpensively using sawdust as the substrate The consumption of oyster mushroom is largely because of its taste and nutritional properties The aims of this research were to reveal the effect of A fumigatus addition in composting process using sawdust substrate for P ostreatus growth medium toward compost quality and yield productivity The experiment revealed that the temperature concentration of glucose xylose and N acetylglucosamine increased and the pH value percentage of cellulose and hemicellulose decreased during 7 days of composting process Based on that biochemical parameter addition of A fumigatus during composting process increased sawdust compost quality Mycelia growth rate per day in treatment group 1 10 cm was faster than control group 1 70 cm during 24 days of observation Statistical test analysis using ANAVA for the fresh weight of P ostreatus indicated that the result was significantly different 0 05 but had no significantly different in diameter of the cap Addition of A fumigatus in sawdust composting process increased yield productivity of P ostreatus ]"

"Human Epidermal Growth Factor (hEGF) merupakan polipeptida yang terdiri atas 53 asam amino. Protein hEGF berfungsi untuk proliferasi sel epitel dan epidermis secara in vitro dan in vivo. Protein hEGF dikode oleh gen EGF. Gen EGFsyn telah dikonstruksi secara sintetik untuk optimasi kodon pada Escherichia coli. Optimasi kodon berfungsi untuk meningkatkan ekspresi protein rekombinan pada E. coli. Subkloning gen EGFsyn ke plasmid pJ404 yang mengandung gen peptida sinyal endoxylanase sangat diperlukan dalam ekspresi protein hEGF rekombinan pada E. coli. Kendala ekspresi protein rekombinan pada E. coli yaitu terbentuknya badan inklusi di sitoplasma. Peptida sinyal endoxylanase digunakan untuk mentranslokasi protein ke periplasma. Digesti gen EGFsyn dan vektor pJ404 dengan enzim NheI dan BamHI diperlukan untuk persiapan subkloning dan ekspresi protein hEGF ke periplasma. Hasil penelitian menunjukkan bahwa gen EGFsyn dan plasmid pJ404 berhasil dipotong dan siap digunakan untuk subkloning.

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Human Epidermal Growth Factor (hEGF) is a polypeptide consisted of 53 amino acids. Its function is to promote epithel and epidermis cells proliferation in vitro and in vivo. It is encoded by EGF gene. An EGFsyn has been constructed to optimize codon usage in Escherichia coli. Codon optimization enhances recombinant protein expression in E. coli. Subcloning EGFsyn gene to a plasmid carrying endoxylanase signal peptide gene is important for recombinant hEGF expression in E. coli. One of the problem expressing recombinant protein in E. coli is the accumulation of inclusion bodies in cytoplasm. Endoxylanase signal peptide is used to translocate protein to periplasm. Digestion of EGFsyn gene and plasmid pJ404 by enzyme NheI and BamHI is needed for preparation of subcloning and hEGF expression to periplasm. The results showed that EGFsyn gene and plasmid pJ404 was digested and can be used for subcloning."

"Ion Magnesium (Mg2+) merupakan salah satu kation divalen yang berperan dalam kondensasi kromosom. Penelitian mengenai pengaruh Mg2+ terhadap kondensasi kromosom dengan menggunakan kromosom yang berasal dari berbagai sel, terutama sel kanker, telah dilakukan. Namun, hingga saat ini, belum ada penelitian yang menggunakan kromosom yang berasal dari sel normal (non-kanker). Salah satu sel yang dapat digunakan adalah limfosit dari sel darah. Penelitian ini bertujuan untuk mengevaluasi pengaruh Mg2+ terhadap struktur kromosom limfosit dengan menggunakan teknik GTL-banding dan karyotyping. Kromosom pada tahap metafase diperoleh dengan mengultur sel darah dan dilanjutkan dengan banding menggunakan Leishman stain. Pengaruh Mg2+ diteliti dengan memberikan larutan XBE5 yang mengandung 5 mM Mg2+ sebagai kontrol, XBE yang mengandung 0 mM Mg2+, dan 1 mM EDTA sebagai chelator kation. Kromosom pada masing-masing perlakuan diberikan penilaian berdasarkan Quality Assesment yang disusun oleh Association for Clinical Cytogenetics berdasarkan International System for Human Cytogenetics Nomenclature (ISCN). Hasil yang diperoleh menunjukkan bahwa kromosom kontrol yang mengandung 5 mM Mg2+ memiliki struktur padat dengan pola banding yang jelas dan intensitas band tebal, sedangkan kromosom pada kelompok perlakuan XBE (0 mM Mg2+) dan 1 mM EDTA memiliki struktur tidak padat dan fibrous dengan pola banding kurang jelas dan intensitas band yang tipis. Nilai rata-rata±SD kromosom kelompok XBE (4,389±0,607) dan EDTA (4,611±0,607) lebih tinggi dibandingkan kontrol XBE5 (4,222±0,427). Hal ini menunjukkan bahwa Mg2+ memiliki pengaruh terhadap struktur kromosom limfosit manusia.

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Magnesium ion (Mg2+ ) is one of the divalent cations that have an important role in chromosome condensation. Researches about the role of Mg2+ have been conducted on the chromosome from various type of cells, especially human cancer cell line. However, there has not been any research conducted using human normal cells such as blood cells lymphocyte chromosome. This study aims to evaluate the effects of Mg2+ on chromosome structure from lymphocyte using GTL-banding and karyotyping. Metaphase chromosomes were obtained by culturing blood cells, followed by banding using Leishman stain. Magnesium ion role was assessed by using XBE5 solution that contains 5 mM Mg2+ as control, XBE solution that contains 0 mM Mg2+, and 1 mM EDTA solution as a cation chelator. The chromosome value from each treatment was evaluated using Quality Assessment from Association for Clinical Cytogenetics according to the International System for Human Cytogenetics Nomenclature (ISCN). The result shows that the control chromosome (XBE5) had a compact structure with a clear banding pattern and bold band intensity, while those treated with XBE and EDTA had a less compact and fibrous structure with an unclear banding pattern and thin band intensity. In addition, the average chromosome score±SD of the XBE (4.389±0.607) and EDTA-treated (4.611±0.607) chromosomes were higher than those of control (4.222±0.427). These data indicated that Mg2+ affects the structure of the human lymphocyte chromosome."

"ABSTRAK Air rebusan jahe telah berhasil digunakan sebagai reduktor untuk biosintesis nanopartikel perak. Biosintesis dilakukan dengan mencampurkan air rebusan rimpang jahe dan larutan AgNO3 yang kemudian diinkubasi selama 24 jam. Karakterisasi larutan hasil reaksi dilakukan dengan fotografi warna larutan dan spektroskopi UV-Vis. Variabel preparasi yang diteliti adalah varietas jahe, fase pertumbuhan, preparasi simplisia, dan perlakuan mekanik. Metode ini digunakan untuk meneliti pengaruh variabel preparasi terhadap nanopartikel perak yang dihasilkan. Varietas jahe yang diteliti adalah jahe gajah, jahe merah, dan jahe emprit. Fase pertumbuhan rimpang yang diteliti adalah bagian anakan dan indukan rimpang. Pengaruh metode preparasi simplisia yang yang diteliti adalah efek bentuk rimpang berupa bubuk dan irisan. Perlakuan mekanik pada larutan saat biosintesis dibedakan antara tanpa pengadukan dan dengan pengadukan. Kemudian diteliti juga pengaruh asam askorbat pada reaksi pembentukan nanopartikel perak. Hasil fotografi menunjukkan bahwa larutan berubah warna dari bening ke kuning kecokelatan yang sesuai dengan warna larutan nanopartikel perak. Spektrum UV-Vis larutan mempunyai nilai absorbansi di panjang gelombang sekitar 420 nm yang bertepatan dengan nilai panjang gelombang absorbansi nanopartikel perak, dengan demikian rimpang jahe dapat digunakan sebagai reduktor biosintesis nanopartikel perak. Di antara varietas jahe, jahe gajah menghasilkan nanopartikel perak yang paling baik, yaitu mempunyai nilai absorbansi 2,2 ± 0,4. Berdasarkan hasil karakterisasi, variabel preparasi yang baik di eksperimen ini adalah penggunaan anakan rimpang sebagai bahan dasar simplisia. Metode preparasi dengan irisan rimpang lebih baik daripada dengan menggiling rimpang, dan perlakuan tanpa pengadukan larutan selama proses reaksi menghasilkan kualitas nanopartikel perak yang lebih baik. Penambahan asam askorbat saat reaksi dapat memperbanyak nanopartikel perak yang dihasilkan.

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ABSTRACT Infution water of ginger has been successfully used as a reductant for biosynthesis of silver nanoparticles. Biosynthesis made ​​by mixing infution water of ginger rhizome and AgNO3 solution then incubated for 24 hours. Characterization of the resulting solution is performed in the color photography solution and UV-Vis spectroscopy. Preparation variables studied were varieties of ginger: gajah ginger, red ginger, and emprit ginger, growth phase: tillers and main rhizomes, form of botanicals material rhizomes: powder and slices, mechanical treatment: mixture solution without stirring and with stirring, and addition of ascorbic acid. The results showed that the photographic color solution changes from clear to yellow brownish that matches the color solution of the silver nanoparticles. UV-Vis spectrum of the solution has a absorbance value at about 420 nm wavelength which coincides with the wavelength of the absorbance value of silver nanoparticles, thus the ginger rhizome can be used as a reductant for biosynthesis of silver nanoparticles. Among the varieties of ginger, gajah ginger produce silver nanoparticles which were the best, which has an absorbance value of 2.2 ± 0.4. Based on characterization results, good preparation variable in this experiment is the use of the tiller rhizomes as the botanicals material rhizomes. Preparation botanicals material rhizomes by slicing ​​better than by grinding, and treatment without stirring the solution during the reaction produce quality of silver nanoparticles better. The addition of ascorbic acid can increase the silver nanoparticles product."

"ABSTRAKGen NS1 merupakan gen penyandi protein NS1 (non-strukural 1) yang terdapat pada virus dengue. Protein NS1 diketahui memiliki potensi untuk dikembangkan sebagai bahan dasar kit diagnostik untuk penyakit demam berdarah dengue (DBD). Penelitian ini bertujuan untuk memperoleh protein rekombinan NS1 virus dengue untuk pengembangan kit diagnostik NS1. Penelitian ini meliputi proses kloning gen NS1 pada vektor ekspresi pYES2/CT, ekspresi pada Saccharomyces cerevisiae INVSc1 dan purifikasi protein rekombinan NS1 menggunakan HisPurTM Ni-NTA Magnetic Beads. Hasil penelitian menunjukkan sebanyak 485 koloni transforman hasil kloning ke dalam E. coli TOP10F? berhasil diseleksi pada medium yang mengandung 100 µg/ml. Analisis hasil PCR dan sekuensing menunjukkan bahwa gen NS1 yang berukuran 1056 pb berhasil terintegrasi ke dalam vektor ekspresi pYES2/CT. Analisis hasil SDS PAGE dan western blotting menunjukkan protein rekombinan NS1 berhasil diekspresikan pada Saccharomyces cerevisiae dengan ukuran sekitar 42?55 kDa. Analisis SDS PAGE untuk hasil purifikasi menunjukkan didapatkan protein yang terelusi dalam kondisi native dengan ukuran sekitar 42?55 kDa. Gen NS1 telah berhasil dikloning dan protein NS1 berhasil terekspresi serta terpurifikasi.

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ABSTRAKNS1 gene is a gene encoding NS1 (non-structural 1) protein dengue virus. Dengue NS1 protein (non-structural 1) is known as an important biomarker for early diagnosis of dengue hemorrhagic fever (DHF) disease. The research objective is to obtain NS1 recombinant protein dengue virus serotype 3 for development kit diagnostic NS1. Stages of the research include cloning NS1 gene Into pYES2/CT expression vector, expression in Saccharomyces cerevisiae INVSc1, and purification NS1 recombinant protein with HisPurTM Ni-NTA Magnetic Beads. A total of 485 colony transformants were selected on medium with ampicilin 100 µg/ml as cloning results in E. coli TOP 10F?. PCR and sequencing analysis showed that NS1 gene was successfully fused to vector pYES2/CT and showed NS1 size is 1056 bp. SDS PAGE and western blotting analysis showed a band of NS1 recombinant protein as expression results. Molecular weight of NS1 protein was approximately 42--55 kDa. SDS PAGE analysis showed a band of NS1 recombinant protein purified in native condition with a molecular weight approximately 42--55 kDa. NS1 gene was successfully cloned, can be also expressed and purified as protein NS1."

"ABSTRAKSubunit protein NS2B-NS3 merupakan protein nonstruktural penyusun virus dengue. Kedua protein tersebut berperan dalam proses replikasi, memodulasi patogenesis, serta respons terhadap sel inang. Penelitian bertujuan untuk memvalidasi hasil kloning yang telah dilakukan oleh peneliti BPPT, mengekspresi dan mempurifikasi protein rekombinan NS2B-NS3 DENV serotipe 3. Vektor yang digunakan untuk kloning dan ekspresi ialah vektor plasmid pYES2/CT. Proses kloning yang telah dilakukan belum tervalidasi sehingga perlu divalidasi dengan metode digesti menggunakan enzim restriksi serta diamplifikasi menggunakan metode PCR. Plasmid yang telah tervalidasi ditransformasi menggunakan metode heat shock ke Saccharomyces cerevisiae sehingga memudahkan proses ekspresi. Hasil ekspresi kemudian divisualisasi menggunakan SDS-PAGE dan western blot. Hasil purifikasi kemudian divisualisasi menggunakan SDS-PAGE saja. Plasmid rekombinan pYES2/CT(NS2B-NS3) yang telah tervalidasi, terekspresi dan terpurifikasi kemudian dikirim untuk proses sekuensing. Hasil visualisasi ekspresi menggunakan metode SDS-PAGE dan western blot ialah terlihat pita spesifik pada ukuran 83 kDa. Hasil visualisasi purifikasi menggunakan SDS-PAGE terlihat muncul pita spesifik pada ukuran 83 kDa pada bagian flow-through dan resin. Hasil sekuensing menunjukan nilai kemiripan 96--99% antara plasmid rekombinan NS2B-NS3 dengan DENV serotipe 3 (DENV-3). Analisis homologi hasil sekuensing dengan isolat nomor 141 asal Jakarta menunjukan nilai 91--94% dan 97% untuk asam amino penyusun DENV.

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ABSTRACTNS2B-NS3 protein subunit are nonstructural protein which construct dengue virus. Both of these proteins take a role in the replication process, modulating pathogenesis, and responding to the host cell. This research aimed to validate the cloning product that had been conducted by BPPT researchers, express and purify recombinant proteins NS2B-NS3 DENV serotypes 3. The vector used for cloning and expression was a plasmid pYES2/CT vector. Furthermore, the cloning product was validated using restriction enzyme digest and amplified using PCR method. Then the plasmid vectors that had been validated were transformed using a heat shock method into Saccharomyces cerevisiae to facilitate the expression process. The results of expression were visualized using SDS-PAGE and western blot. Whereas, the results of purification were visualized using SDS-PAGE only. Recombinant plasmid pYES2/CT (NS2B-NS3) that had been validated, expressed, and purified were proceed to the sequencing process. The visualized expression showed a band of 83 kDa. The visualized purification showed a band of 83 kDa in the flow-through and resin part. The sequencing results showed 96--99% sequence similarity between NS2B-NS3 recombinant plasmid and DENV serotypes 3 (DENV-3). The homology analysis of the sequencing results using isolates number 141 from Jakarta showed 91--94% value and 97% for the amino acid which construct DENV."