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The effect of DMSO on ITS2 amplification in the molecular identification of Anopheles farauti Laveran (Diptera: Culicidae), from a colony established in the laboratory Suyono, Ign. Joko; Situmorang, Jesmandt; Soesilohadi, R.C. Hidayat; Pratiwi, Rarastoeti
Jurnal Entomologi Indonesia Vol 10, No 2 (2013): September
Publisher : Perhimpunan Entomologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (429.779 KB) | DOI: 10.5994/jei.10.2.93

Abstract

Identifikasi spesies sibling sangat penting untuk mempelajari epidemiologi malaria. Ciri morfologi secara umum digunakan dalam identifikasi spesies Anopheles, namun tidak sesuai ketika digunakan untuk spesies sibling dan spesies kriptik. Analisis dengan menggunakan PCR-RFLP dari ITS2 rDNA pada saat ini merupakan metode yang paling cocok dan sensitif untuk membedakan spesies Anopheles punctulatus group. Tujuan penelitian ini adalah untuk mengetahui pengaruh konsentrasi DMSO terhadap amplifikasi ITS2 dalam mengidentifikasi An. farauti dari koloni di BATAN Jakarta dengan PCR-RFLP berdasarkan ITS2 rDNA. Hasil penelitian menunjukkan bahwa penambahan DMSO dengan konsentrasi 6% dan 7% menghasilkan produk amplifikasi ITS2 dengan ukuran 750 pb. DMSO dapat digunakan dalam PCR untuk mencegah terbentuknya struktur sekunder ketika mengamplifikasi template dengan kandungan GC yang tinggi. Identifikasi molekular An. farauti dengan menggunakan PCR-RFLP merupakan Anopheles farauti sensu stricto.
MUTASI MISSENSE (P.374PHE/LEU) PADA EKSON 5 GEN MATP, PENYEBAB OCULOCUTANEOUS ALBINISM TIPE 4 (OCA4) DI WONOSOBO, JAWA TENGAH Nur Handayani, Niken Satuti; Sukmawati, Feri; Pratiwi, Rarastoeti
Prosiding Seminar Biologi Vol 8, No 1 (2011): Seminar Nasional VIII Pendidikan Biologi
Publisher : Prodi Pendidikan Biologi FKIP UNS

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Abstract

ABSTRAK Albinisme merupakan kelainan genetik autosomal resesif  berupa gangguan sintesis melanin yang terjadi pada manusia. Albinisme dibagi menjadi dua kelompok besar, yaitu Ocular Albinism (OA) dan Oculocutaneous Albinism (OCA). Berdasarkan gen yang mengalami mutasi, OCA dibedakan menjadi 4 tipe yaitu OCA1, OCA2, OCA3 dan OCA4. OCA4 disebabkan mutasi pada gen MATP. Penelitian yang telah dilakukan dengan PCR-SSCP (Polymerase Chain Reaction-Single Stranded Conformation Polymorphism) mendeteksi adanya mutasi pada ekson 5 gen MATP, pada penderita albinisme di Wonosobo, Jawa Tengah. Sekuensing ekson 5 gen MATP dilakukan untuk mengidentifikasi tipe mutasinya. DNA diisolasi dari sampel darah penderita dan digunakan sebagai template untuk amplifikasi ekson 5 gen MATP dengan metode PCR. Produk PCR selanjutnya digunakan sebagai template untuk sekuensing dengan metode Sanger. Hasil sekuensing dianalisis menggunakan program Clustal-W dan dibandingkan dengan sekuens ekson 5 gen MATP dari International DNA Data Base (nomer akses AF172849.1). Hasil analisis menunjukkan adanya pada perubahan basa nukleotida no.1122 dari C menjadi G (c.1122 C>G) yang mengakibatkan mutasi missense, yaitu fenilalanin menjadi leusin, pada asam amino nomer 374 (p.374 Phe/Leu).   Kata kunci : OCA4, mutasi, ekson 5, gen MATP
KADAR SERUM GLUTAMIC PYRUVATE TRANSAMINASE DARAH TIKUS (Rattus norvegicus Berkenhout, 1769) HIPERLIPIDEMIA DENGAN ASUPAN PELET NASI DAN BEKATUL BERAS HITAM PADI (Oryza sativa L.) “CEMPO IRENG” Chasanah, Siti Nur; Pratiwi, Rarastoeti
CENDEKIA EKSAKTA Vol 4, No 1 (2019)
Publisher : Universitas Wahid Hasyim

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.3194/ce.v4i1.2672

Abstract

Diet makanan tinggi lemak jenuh dan tinggi kolesterol dapat meningkatkan resiko terjadinya kondisi hiperlipidemia. Penelitian sebelumnya, menunjukkan bahwa beras merah dan hitam mengandung antioksidan yang lebih tinggi dibandingkan beras putih. Beras hitam efektif dalam memperbaiki fungsi hati tikus yang mengalami hiperlipidemia. Penelitian ini bertujuan untuk mengetahui kadarSerum Glutamic Pyruvate Transaminase (SGPT) pada tikus hiperlipidemia dengan perlakuan diet nasi hitam dan bekatul beras hitam yang berasal dari padi hitam ?Cempo Ireng?. Sebanyak 24 ekor tikus putih jantan yang dibagi menjadi 4 kelompok yaitu kelompok kontrol normal dan 3 kelompok hiperlipidemia yang masing-masing kelompok diberi diet pelet pakan basal, pelet mengandung 30% nasi hitam dan pelet mengandung 10% bekatul beras hitam. Tahap pertama adalah induksi hiperlipidemia pada kelompok hiperlipidemia dengan pemberian campuran minyak babi dan kuning telur bebek dengan perbandingan 1:1 (v/v) yang dicampur dengan kolesterol murni 2%. Dosis yang diberikan sebanyak 1 ml/100 g BB secara peroral setiap hari selama 30 hari. Tahap kedua adalah perlakuan diet pakan selama 30 hari dengan tetap mempertahankan kondisi hiperlipidemia. Berat badan tikus ditimbang setiap 7 hari dan sisa konsumsi pakan ditimbang setiap hari. Pengukuran kadar SGPT dilakukan setelah perlakuan hiperlipidemia dan perlakuan diet pakan. Hasil pengukuran kadar SGPT dianalisis menggunakan one way ANOVA dilanjutkan uji Tukey?s HSD dan uji T-Test dengan signifikansi 5%. Hasil analisis menunjukan bahwa saat perlakuan hiperlipidemia disertai pakan mengandung 30% nasi hitam mampu menurunkan kadar SGPT sebanyak 19,38%. Pada perlakuan hiperlipidemia disertai pakan pelet mengandung 10% bekatul beras hitam mampu menurunkan kadar SGPT sebanyak 21,46%. Secara statistik penurunan ini signifikan (<0,05), sehingga dapat disimpulkan bahwa bekatul beras hitam lebih efektif dari pada nasi hitam dalam memperbaiki fungsi hati berdasarkan penurunan kadar SGPT pada tikus hiperlipidemia. Kata kunci: SGPT, hiperlipidemia, beras hitam, bekatul 
Cloning and Expression of ORF124 Koi Herpesvirus as a Vaccine ., Murwantoko; Pratiwi, Rarastoeti; Kawaichi, Masashi
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Abstract

Koi herpesvirus (KHV) which also known as Cyprinid herpesvirus 3 (CyHV-3), Koi herpes-like virus, and carp interstitial nephritis gill necrosis virus (CNGV), caused signifi cant morbidity and mortality in koi and common carp (Cyprinus carpio). The case fatality rate of this disease is 80&ndash;100%. Glycoprotein has been used for vaccine development as sub unit vaccine against viruses. The aim of this research was to clone and express membrane glycoprotein ORF124 KHV as a candidate of recombinant vaccine. ORF124 KHV gene was successfully cloned into pBSKS and sequenced. Result showed that ORF124 KHV (isolate from Indonesia) had 100 % similarity with Cyprinid herpesvirus 3 strain TUMST1 (from Japan), 99% similarity with Koi herpesvirus strain KHV-U (from USA) and Koi herpesvirus strain KHV-I (from Israel). Prediction analysis of T and B cell epitopes showed that ORF124 KHV protein had 14 and 11 T cell epitopes (IAd, Rothbard/Taylor pattern),and had 10 B cell epitopes, suggested that the protein can be used as a vaccine candidate. ORF124 gene has been expressed in Escherichia coli under pET32-a(+)vector.
Cloning and Sequence Analysis of Capsid Protein Gene of Iridovirus Indonesian Isolates ., Murwantoko; Handayani, Christina Retna; Pratiwi, Rarastoeti
Indonesian Journal of Biotechnology Vol 14, No 1 (2009)
Publisher : Universitas Gadjah Mada

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Abstract

Iridovirus was known as agents that caused serious systemic disease in freshwater and marine fishes. The mortality up to 100% of orange-spotted grouper (Epinephelus coioides) due to iridovirus infection has been reported in Indonesia. The gene encoding capsid protein of iridovirus is supposed to be conserved and has the potency for the development of control methods. The objectives of this study are to clone the gene encoding capsid protein iridovirus and to analyze their sequences. The&nbsp;&nbsp; spleen tissues of orange-spotted grouper were collected and extracted their DNA. The DNA fragment of capsid protein of iridovirus genes were amplified by PCR using designed primers with the extraction DNA as templates. The amplified DNA fragments were cloned in pBSKSII and sequenced.&nbsp; The genes encoding capsid protein of iridovirus from Jepara and Bali were successfully amplified and cloned. The Jepara clone (IJP03) contained complete open reading frame (ORF) of the gene composed by 1362 bp nucleotides which encoded 453 amino acids. Those Jepara and Bali (IGD01) clones shared 99.8% similarity in nucleotide level and 99.4% at amino acid level. Based on those sequences, Indonesian iridovirus was belonged to genus Megalocystivirus and shared 99,6-99,9% similarity on nucleotide level with DGIV, ISKNV, MCIV, and ALIV
Characterization of envelope-transmembrane Gene of Jembrana Disease Virus Tabanan 1995 Isolate Kusumawati, Asmarani; Pratiwi, Rarastoeti; Astuti, Pudji; Hamid, Penny Humaidah
Indonesian Journal of Biotechnology Vol 15, No 1 (2010)
Publisher : Universitas Gadjah Mada

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Abstract

The availability of specific and rapid detection methods is essential for monitoring the health status of farmed species, particularly in viral disease as in this case early diagnosis is a critical factor in containing disease outbreaks. Jembrana Disease Virus (JDV) is a lentivirus that causes an acute, severe disease syndrome in infected Bali cattle in Indonesia, resulting in heavy economic losses because of the high mortalities. The virus-host interaction and the modes of transmission are still unknown. The goal of the research was to designa probe candidate of Jembrana Disease Virus based on envelope-transmembrane (env-tm) gene to optimize Jembrana disease detection method. The DNA fragment derived from env-tm of JDV was used, cloned in pGEX-TM and expressed in E.coli DH 5&alpha;. Sequence analysis was conducted with BLAST programs from NCBI. Sequence analyses of the fragments of env-tm clone, indicated that it has a very closed genetic relation with 97,68% homology identity. Probe was designed based on the conserved region of env-tm using Geneious resulted in JT2 252 bp long. BLAST analyses showed that probes had high specifity to other strains of JDV in Indonesia.Key words : probe, env-tm, JDV, specifity, sensitivity.
The effect of ethanolic extract of black and white rice bran (Oryza sativa L.) on cancer cells Rukmana, Rizal Maarif; Soesilo, Nyoman Puniawati; Rumiyati, Rumiyati; Pratiwi, Rarastoeti
Indonesian Journal of Biotechnology Vol 21, No 1 (2016)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1157.736 KB) | DOI: 10.22146/ijbiotech.26814

Abstract

Indonesia has a wide range of rice cultivars and pigments. This rice can be used as a source of phytochemical compounds for cancer prevention. This research aims to analyze the cytotoxic activities of the ethanolic extract of black rice bran of 4 local cultivars i.e. ‘Cempo Ireng’, ‘Woja Laka’, ‘Toraja’ and ‘IR­64’ (white rice) on  cancer cells and to determine the compounds groups of those extracts. First step, rice bran was extracted with ethanol. This extract was applied to Raji (a human Burkitt Lymphoma cancer), HepG2 (a human liver cancer), and Vero (a nonhuman cell line) cells in order to measure the cytotoxic activities by using MTT assay. To determine descriptively the compounds groups of phenolics, flavonoids, terpenoids, steroids, and alkaloids the thin layer chromatography method was performed. The IC50 value was analyzed quantitatively by using probit analysis. Results showed that the IC50 values of ethanolic extract of rice bran ‘Woja Laka’, ‘Toraja’, ‘Cempo Ireng’ and ‘IR 64’ on HepG2 cells were 857.23±99.19; 1,896.55±83,8; 1,494.47±87.81 and 727.89±145,97 µg/ml respectively. The IC50 on Raji cells were 816.61±85.31; 1,079.93±28.31; 1,627.82; ±119.82, and 769.33±61.43 µg/ml respectively. The IC50 on Vero cells were 1,295.2±37; 1,232.07±165.51; 1,874.14±169.56, and 724.4±122.79 µg/ml respectively. The ethanolic extracts of rice bran from four cultivars contain phenolics, flavonoids, terpenoids, and steroids. However, alkaloids could not be detected. The variety of rice cultivars indicates the variation of cytotoxic activities on cancer cells. The ethanolic extracts of rice bran from those four rice cultivars contain similar kinds of organic compounds groups but vary in the Rf values.
Pengaruh Perbedaan Waktu Awal Pemberian Pakan Buatan terhadap Pertumbuhan dan Aktivitas Enzim Pencernaan Larva Ikan Kerapu Bebek Cromileptes altivelis (Valenciennes 1828) Melianawati, Regina; Pratiwi, Rarastoeti; Astuti, Ni Wayan Widya
JURNAL BIOLOGI INDONESIA Vol 8, No 2 (2012): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v8i2.3055

Abstract

The effect of different initial time of artificial feeding to the growth and the activities of digestive enzymes of humpback grouper larvae Cromileptes altivelis (Valenciennes 1828).Humpback grouper is an economically marine finfish commodity, so it is important to scale up its culture. In aquaculture, feed is dominantly affecting to fish growth. Besides live feeds, theartificial feed is also given to fulfill the need of nutrient for larvae. However, the capability of larvae to digest the artificial feed is still limited because of its simplicity and immaturity of digestive system structure. Digestive enzymes are a biological indicator to figure out the capability of larvae to digest the artificial feed. This research was conducted to find out the influence of different initial time of artificial feeding to the growth and digestive enzyme activity of humpback grouper larvae. Larvae rearing were done in hatchery. Larvae were fed with live feed rotifers started in the evening of the second day. The treatment given in this research was the difference initial time of artificial feeding, i.e: (A) given at 8 days old larvae and (B) given at13 days old larvae. The variable observations were the growth and digestive enzyme activity of larvae. The results indicate that the different initial time of artificial feeding influenced to thegrowth and digestive enzyme activity of humpback grouper larvae. Total length, body weight, length of dorsal spine and ventral spine of 30 days old humpback grouper larvae on treatment A were 17.47±2.35 mm, 0.11±0.04 g, 6.83±0.45 mm and 5.07±0.50 mm, respectively, while on treatment B were 13.23±2.53 mm, 0.04±0.02 g, 5.73±1.11 mm and 4.48±0.50 mm, respectively. Up to 30 days old larvae, protease and amylase enzymes activities of larvae on treatment A was higher than on treatment B, while lipase enzymes activity of larvae on treatment B was higher than on treatment A. Therefore, the different initial time of artificial feeding gives the different effect on the growth and the activities of protease, amylase and lipase enzymes of humpback grouper larvae.Key words: artificial feed, growth, digestive enzyme activity, humpback grouper, larvae
DETEKSI GAMMA AMINO BUTYRIC ACID (GABA) PADA DAUN Artocarpus altilis Indrowati, Meti; Astuti, Pudji; Pratiwi, Rarastoeti; Rumiyati, Rumiyati
Prosiding SNPS (Seminar Nasional Pendidikan Sains) 2015: Prosiding SNPS (Seminar Nasional Pendidikan Sains)
Publisher : Universitas Sebelas Maret Surakarta

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Abstract

Senyawa aktif Gamma Amino Butyric Acid (GABA) merupakan asam amino yang terdapat di otak mammalia dan berperan sebagai neurotransmiter.  Selain pada sistem saraf pusat, GABA juga terdapat dalam konsentrasi tinggi di sel β pankreas serta memiliki potensi sebagai antidiabetes. GABA dapat disintesis oleh hewan, bakteri asam laktat dan tumbuhan.  Penelitian dilakukan dengan tujuan mendeteksi keberadaan GABA dalam bagian tumbuhan yaitu daun Artocarpus altilis.  Deteksi dilakukan dengan Kromatografi Lapis Tipis (KLT) menggunakan fase diam plat silika, fase gerak BAW(3:1:1), dan GABA murni sebagai pembanding.  Larutan GABA dibuat dengan pelarut etanol:air (7:3).  Hasil penelitian menunjukkan GABA terdeteksi pada ekstrak daun Artocarpus altilis.
Pemanfaatan Limbah Buah Salak Pondoh Sebagai Substrat Nata De Salacca Melalui Aplikasi Bioteknologi di Dusun Tegal Domban, Sleman, Yogyakarta Pratiwi, Rarastoeti; Lestari, Fajar Budi; Widianto, Donny
Jurnal Pengabdian kepada Masyarakat (Indonesian Journal of Community Engagement) Vol 1, No 1 (2015): September
Publisher : Direktorat Pengabdian kepada Masyarakat Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpkm.16937

Abstract

Dusun Tegal Domban is one of the highest Salak Pondoh produce in Sleman District. However there is a problemfaced by salak farmers due to the overproduction and overripe which mightcause waste. Implementation of Biotechnology should be one of the alternativesolutions to overcome this problem. The aims of the program were to utilize the salak fruits wasteto produce Natade Salaccausing biological agents, such as Acetobacter xylinum, and to implement the education for sustainability development which subjected toa women organization PKK Dusun Tegal Domban who accompanyed by the team from Biotechnology Master Program UGM in order to learn how to produce Nata de Salacca. This program was initiated with the laboratory experiments to find the fine composotion of nata substrates to get the optimal nata product. The following action was to acompany the women who aplicate the nata production process. Results from the laboratory experiment showed that the best composition of substrat and water ratiowas 1:4, and the nata thickness was 0.62 cm, while the nata weight was 542.22 g. Those parameters were used for nata standard indicators.Results from the activities of nata production by women groups PKK Dusun Tegal Domban were showed the similar results with the nata standard for thickness, however nata weight was slightly lower than the nata standard.