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Effect of Culture Medium Supplementation with b-mercaptoethanol and Amino Acid on Canine Intergeneric Embryo Development with Porcine Oocyte Cytoplasm Recipient Fibrianto, Yuda Heru
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

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The present study investigated the effect of culture medium supplementation with mercaptoethanol ( ME)and amino acid (AA) on canine intergeneric embryo development with porcine oocyte cytoplasm. Porcine cumulusoocyte complexes (COCs) were collected from slaughterhouse and matured in TCM-199 supplemented with 26.2mM NaHCO, 3.05 mM glucose, 0.91 mM sodium pyruvate, 0.57 mM L-cysteine, 75 mg/l kanamycin, 10 ng/ml 3epidermal growth factor, equine chorionic gonadotropin (eCG), 10 IU/ml human chorionic gonadotropin (hCG),and 10% (v/v) porcine follicular fluid (pFF) at 39 °C in a humidified atmosphere of 5% CO for 42-44 h and donor cell 2collected from ear skin afghanhound male dog. After somatic cell nuclear transfer (SCNT), embryo developmentwere examined for cleavage rate and 144 hr for final development after cultured in media. The result shows that,amino acid and mercapoethanol addition in culture medium (NCSU-23) have no effect on embryo development.The development rate of embryo until 16 cell stage in NCSU and NCSU supplement are 4.67% and morula stage are3.73% and 4.67%.Key words : intergeneric clone embryo, canine, ( amino acid (AA)

Genetic Heterogenity Profile of Penaeus monodon Broodstock F1 Revealed by Mitochondria DNA-RFLP and RAPD Prastowo, Bambang Widyo; Rahardianti, Rahayu; Nur, Evi Maftuti; Taslihan, Arief
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

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The genetic heterogeneity of Penaeus monodon broodstock F1 was evaluated using Restriction Fragment LengthPolymorphism (RFLP-mtDNA) and Random Amplified Polymorphic DNA (RAPD) analysis. The RFLP analysis wasconducted by amplifying 16SrDNA region and digested with restriction enzyme Nde II. According to the RFLPanalysis, heterogeneity value of P. monodon F1 broodstock population is 0,0422; male F1 population is 0,0613 andfemale F1 population is 0,1252. The primer OPA2 was used in RAPD analysis. According to the RAPD analysis,heterogeneity value of P. monodon F1 broodstock population is 0,0417; male F1 population is 0,0653 and female F1population is 0,1104. The results of this research showed that either RFLP or RAPD can be used as a family specificmarker for Penaeus monodon.Key words : Penaeus monodon, RFLP, RAPD, heterogeneity, genetic marker

Biochemival Characterization of an Antibactrial Glycoprotein from Achatina fulica ferussac Snail Mucus Local Isolate and Their Implication on Bacterial Dental Infection Berniyanti, Titiek; Waskito, Edy Bagus; s, Suwarno
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

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Snails crawl over a variety of potentially contaminated surfaces and their foot is the primary site of entry forpathogens, parasites and a range of opportunistic organisms, so it is a little wonder that they must have a defensivesystem to protect them. The mucus secreted on the body surfaces of mollusks is known to play crucial role inlocomotion, feeding, osmoregulation, reproduction and protection of epithelial surfaces. The snail mucus alsocontains Glycoaminoglycans (GAGs) which are complex polysaccharides that participate in the regulation ofphysiological processes through the interactions with a wide variety of proteins. GAGs, such as heparin, serve as keyto biological response modifiers, in example for acting asa a target for pathogen and parasitic factors for attachment,invasion, and immune system.For years, it has been known that the mucus secretions from snails Achatina fulica ferussac local isolate can be usedas a medication, and even empirically it is used to treat infected teeth tahat is suffered by people in rural area. Theantibacterial factor was surveyed in the aqueous extract and the mucin fraction of snail Achatina fulica ferussac, andthey exhibited positive antibacterial for Gram-positive, Escherichia coli and Gram negative, Streptococcus mutans. Inthe following study, it has been proved that an antibacterial content in the mucus was a Glycoprotein. It wascomposed of two subunits of Molecular Weight (MW) 71-73 kDa. The GelCode Glycoprotein Staining Kit detectedglycoprotein sugar moieties in polyacrylamide gel and on nitrocellulose membrane, while the glycoproteincarbohydrate estimation kit detected glycoprotein and estimated carbohydrate content. The glycoprotein contentwas 4.537 ± 0.876 for carbohydrate and 6.420 ± 1.242 for protein.Keywords : characterization, glycoprotein, Achatina fullica Ferussac snail mucus, galur Jawa, antibacterialfactor

Alternative Oxidase (AtAox) c78s Mutant Expression at Escherichia coli (SASX41DB) Djajanegara, Ira
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

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Alternative oxidase (AOX) is the terminal oxidase operating in the mitochondrial electron transport chain. Theenzyme is activated by organic acid such as pyruvate and by reduction process. Based on sequences alignment ofalternative oxidase gene (Aox) found in several organisms, there are 2 conserved cysteine residues. In order toinvestigate the importance of those cysteine residues on the activity of AOX, mutation at cysteine residue number 78of Aox gene isolated from Arabidopsis thaliana (AtAox) was conducted. Cysteine at position number 78 was changedinto serine and the c78s mutant was expressed in Escherichia coli strain SASX41DB. This particular E. coli strain isunable to grow aerobically unless transformed with Arabidopsis Aox gene (AtAox). Expression studies on c78smutant showed that this mutant cannot be oxididized and can not be activated by pyruvic acid. This mutant isacivated by succinate instead of pyruvate. Mutation at cysteine closer to the N residue is affecting both organic acidand redox activation. Therefore, it is concluded that cysteine residue closer to the N residue is the site for bothactivation by pyruvate as well as activation by reduction process.Keywords : Alternative oxidase, site-directed mutation, SASx41DB, cysteine residues

Effect of Staurosporine on the Intracellular Localization of Hepatitis B Virus Core Protein Haryanto, Aris; Wijayanti, Nastiti; Kann, Michael
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

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protein is also including in the HBV genome targeting into the nucleus through modulating carboxyl residues byphosphorylation. Nuclear localication Signal (NLS) in HBV core protein is inside the virion structure and it must beunmasked in order to function, perhaps by phosphorylation. Phosphorylation of of HBV core protein in turn couldbegin to alter capsid conformation. Staurosporine is a natural product originally isolated from bacteriumStreptomyces staurosporeus. Staurosporine was discovered to have biological activities ranging from anti-fungal toanti-hypertensive. The interest in these activities resulted in a large investigative effort in chemistry and biology andthe discovery of the potential for anti-cancer treatment. The main biological activity of Staurosporine is the inhibitionof protein kinases through the prevention of ATP binding to the kinase. In the present study, we have studied theintracellular localization of EGFP-Core fusion protein with triple HBV core and SV-40 nuclear localization signal atits carboxyl terminal in presence and absence of Staurosporine. We also to study the effect of Staurosporine treatmenton the intracellular localization of EGFP-Core fusion protein in the hepatocyte cells line of HepG2 cell. Resultsshowed that effect of Staurosporine is prevent the nuclear localization of EGFP-Core fusion protein into nucleusthrough an inhibition of the phosphorylation of core protein. Stauroporine also prevents cell division so that passivetrapping of core protein is inhibited.

Superoxide Dismutase of Micrococcus sp. S2 and Its Involve in Paraquat Detoxification Margino, Sebastian; Martani, Erni; Magdalena, Medhina
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

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As an active ingredient of herbicide, paraquat will induce formation of superoxide radicals. The previousresearch succeeded in isolating paraquat degrading bacteria from peat soil, Micrococcus sp. S2, that tolerant to highconcentration of paraquat. An anti-oxidative enzyme, namely superoxide dismutase (SOD, EC.1.15.1.1), wasbelieved to be responsible for the paraquat tolerance. This research was conducted to study the characteristic of theSOD synthesize by Micrococcus sp. S2 and its ability on neutralize superoxide which arise from paraquat reoxidation.To observe the effect of paraquat on Micrococcus sp. S2, the bacteria was grown in 10% Luria Bertani brothmedium amended with several concentrations of paraquat, from 0 (control) up to 100 mg/ml. Within incubationtime of 72 hours, bacterial growth, activity of superoxide dismutase and paraquat residue were analyzed. Theisozymes of superoxide dismutase were distinguished using two kinds of specific inhibitor, namely HO and KCN. 2 2The results showed that paraquat significantly inhibit the growth of Micrococcus sp. S2. The higher paraquatcocentration in the medium caused the higher growth inhibition. However, the bacteria is still survive in the mediumcontaining toxic herbicide, and this ability was suggested related to superoxide dismutase activity in removing thesuperoxide radicals. Analysis using gel electrophoresis indicated that at least three types of SOD isozyme weresynthesized by Micrococcus sp. S2; they were Ferri-SOD (Fe-SOD), Mangani-SOD (Mn-SOD), and the last one wassuspected to be the Cupro Zinc-SOD (CuZn-SOD). The Mangani-SOD was suspected to play an important roles ondetoxifying superoxide which arise from paraquat oxidation.Keywords : Micrococcus sp.S2, paraquat, superoxide dismutase, isozymes

Biochemival Characterization of an Antibactrial Glycoprotein from Achatina fulica ferussac Snail Mucus Local Isolate and Their Implication on Bacterial Dental Infection Titiek Berniyanti; Edy Bagus Waskito; S. Suwarno
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

Snails crawl over a variety of potentially contaminated surfaces and their foot is the primary site of entry forpathogens, parasites and a range of opportunistic organisms, so it is a little wonder that they must have a defensivesystem to protect them. The mucus secreted on the body surfaces of mollusks is known to play crucial role inlocomotion, feeding, osmoregulation, reproduction and protection of epithelial surfaces. The snail mucus alsocontains Glycoaminoglycans (GAGs) which are complex polysaccharides that participate in the regulation ofphysiological processes through the interactions with a wide variety of proteins. GAGs, such as heparin, serve as keyto biological response modifiers, in example for acting asa a target for pathogen and parasitic factors for attachment,invasion, and immune system.For years, it has been known that the mucus secretions from snails Achatina fulica ferussac local isolate can be usedas a medication, and even empirically it is used to treat infected teeth tahat is suffered by people in rural area. Theantibacterial factor was surveyed in the aqueous extract and the mucin fraction of snail Achatina fulica ferussac, andthey exhibited positive antibacterial for Gram-positive, Escherichia coli and Gram negative, Streptococcus mutans. Inthe following study, it has been proved that an antibacterial content in the mucus was a Glycoprotein. It wascomposed of two subunits of Molecular Weight (MW) 71-73 kDa. The GelCode Glycoprotein Staining Kit detectedglycoprotein sugar moieties in polyacrylamide gel and on nitrocellulose membrane, while the glycoproteincarbohydrate estimation kit detected glycoprotein and estimated carbohydrate content. The glycoprotein contentwas 4.537 ± 0.876 for carbohydrate and 6.420 ± 1.242 for protein.Keywords : characterization, glycoprotein, Achatina fullica Ferussac snail mucus, galur Jawa, antibacterialfactor

Superoxide Dismutase of Micrococcus sp. S2 and Its Involve in Paraquat Detoxification Sebastian Margino; Erni Martani; Medhina Magdalena
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

As an active ingredient of herbicide, paraquat will induce formation of superoxide radicals. The previousresearch succeeded in isolating paraquat degrading bacteria from peat soil, Micrococcus sp. S2, that tolerant to highconcentration of paraquat. An anti-oxidative enzyme, namely superoxide dismutase (SOD, EC.1.15.1.1), wasbelieved to be responsible for the paraquat tolerance. This research was conducted to study the characteristic of theSOD synthesize by Micrococcus sp. S2 and its ability on neutralize superoxide which arise from paraquat reoxidation.To observe the effect of paraquat on Micrococcus sp. S2, the bacteria was grown in 10% Luria Bertani brothmedium amended with several concentrations of paraquat, from 0 (control) up to 100 mg/ml. Within incubationtime of 72 hours, bacterial growth, activity of superoxide dismutase and paraquat residue were analyzed. Theisozymes of superoxide dismutase were distinguished using two kinds of specific inhibitor, namely HO and KCN. 2 2The results showed that paraquat significantly inhibit the growth of Micrococcus sp. S2. The higher paraquatcocentration in the medium caused the higher growth inhibition. However, the bacteria is still survive in the mediumcontaining toxic herbicide, and this ability was suggested related to superoxide dismutase activity in removing thesuperoxide radicals. Analysis using gel electrophoresis indicated that at least three types of SOD isozyme weresynthesized by Micrococcus sp. S2; they were Ferri-SOD (Fe-SOD), Mangani-SOD (Mn-SOD), and the last one wassuspected to be the Cupro Zinc-SOD (CuZn-SOD). The Mangani-SOD was suspected to play an important roles ondetoxifying superoxide which arise from paraquat oxidation.Keywords : Micrococcus sp.S2, paraquat, superoxide dismutase, isozymes

Genetic Heterogenity Profile of Penaeus monodon Broodstock F1 Revealed by Mitochondria DNA-RFLP and RAPD Bambang Widyo Prastowo; Rahayu Rahardianti; Evi Maftuti Nur; Arief Taslihan
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

The genetic heterogeneity of Penaeus monodon broodstock F1 was evaluated using Restriction Fragment LengthPolymorphism (RFLP-mtDNA) and Random Amplified Polymorphic DNA (RAPD) analysis. The RFLP analysis wasconducted by amplifying 16SrDNA region and digested with restriction enzyme Nde II. According to the RFLPanalysis, heterogeneity value of P. monodon F1 broodstock population is 0,0422; male F1 population is 0,0613 andfemale F1 population is 0,1252. The primer OPA2 was used in RAPD analysis. According to the RAPD analysis,heterogeneity value of P. monodon F1 broodstock population is 0,0417; male F1 population is 0,0653 and female F1population is 0,1104. The results of this research showed that either RFLP or RAPD can be used as a family specificmarker for Penaeus monodon.Key words : Penaeus monodon, RFLP, RAPD, heterogeneity, genetic marker

Effect of Culture Medium Supplementation with b-mercaptoethanol and Amino Acid on Canine Intergeneric Embryo Development with Porcine Oocyte Cytoplasm Recipient Yuda Heru Fibrianto
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

The present study investigated the effect of culture medium supplementation with mercaptoethanol ( ME)and amino acid (AA) on canine intergeneric embryo development with porcine oocyte cytoplasm. Porcine cumulusoocyte complexes (COCs) were collected from slaughterhouse and matured in TCM-199 supplemented with 26.2mM NaHCO, 3.05 mM glucose, 0.91 mM sodium pyruvate, 0.57 mM L-cysteine, 75 mg/l kanamycin, 10 ng/ml 3epidermal growth factor, equine chorionic gonadotropin (eCG), 10 IU/ml human chorionic gonadotropin (hCG),and 10% (v/v) porcine follicular fluid (pFF) at 39 °C in a humidified atmosphere of 5% CO for 42-44 h and donor cell 2collected from ear skin afghanhound male dog. After somatic cell nuclear transfer (SCNT), embryo developmentwere examined for cleavage rate and 144 hr for final development after cultured in media. The result shows that,amino acid and mercapoethanol addition in culture medium (NCSU-23) have no effect on embryo development.The development rate of embryo until 16 cell stage in NCSU and NCSU supplement are 4.67% and morula stage are3.73% and 4.67%.Key words : intergeneric clone embryo, canine, ( amino acid (AA)

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Daerah istimewa yogyakarta

INDONESIA