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Indonesian Journal of Biotechnology

ijbiotech
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Published by Universitas Gadjah Mada

ISSN : 08538654 EISSN : 20892241 DOI : -

Core Subject : Science,

Biochemistry, Genetics & Molecular Biology Immunology & microbiology Materials Science & Nanotechnology

The Indonesian Journal of Biotechnology (IJBiotech) is an open access, peer-reviewed, multidisciplinary journal dedicated to the publication of novel research in all aspects of biotechnology, with particular attention paid to the exploration and development of natural products derived from tropical—and especially Indonesian—biodiversity. IJBiotech is published biannually and accepts original research articles featuring well-designed studies with clearly analyzed and logically interpreted results. A strong preference is given to research that has the potential to make significant contributions to both the field of biotechnology and society in general.

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Articles 518 Documents

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Detection of eae, bfpA, espA Genes on Diarrhoeagenic Strains of Escherichia coli Isolates Agnes Sri Harti; Susi Iravati; Widya Asmara
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada

The Enteropathogenic Escherichia coli (EPEC) is one of pathogenic strain of diarrheagenic E. coli group in children and infant that occurs in developing countries. The significant virulence factors in pathogenic EPEC are eaeA (E. coli attaching- effacing ), bfpA (bundle-forming pilus A) and espA (encoding secreted protein A) genes. The use of DNA probes to detect the virulence genes in E. coli in Indonesia is not common yet. In this experiment the gene fragments of eae, bfpA, and espA were used as probes to detect the EPEC among E. coli isolates from stool specimensin of diarrheic children attending Public Health Centers in Yogyakarta. The DNA samples were isolated from 49 diarrheagenic E. coli isolates. The DNA probes of eae, bfpA and espA were obtained by amplification of DNA fragment of EPEC O126 using PCR technique. Furthermore, those probes were used to identify the presence of those genes among E. coli isolates using hybridization technique. The results showed that 42 (85.7%) isolates were espA+, 25 isolates (51%) were eaeA+ (EPEC strains). Therefore among 25 isolates of EPEC, 20 isolates (80 %) among EPEC were bfpA+ (typical EPEC strains).

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

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.

Production of Poly-α-hydroxybutyrate (PHB) from Sago Starch by The Native Isolate Bacillus megaterium PSA10 Nur Arfa Yanti; Langkah Sembiring; Sebastian Margino
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada

A new bacterial strain that produces amylase and poly-α-hidroxybutyrate (PHB) using sago starch as carbon source was characterized and identified to be member of the Bacillus megaterium group based on phenotypic characteristics and 16S rDNA gene sequences. Amylase activity was determined spectrophotometrically on the basis of substrate concentration reduction. PHB production was quantified with UV spectrophotometer. The polymer produced by B. megaterium PSA10 was identified by Fourier Transform Infrared spectroscopy (FTIR). The result of the study showed that the amylase specific activity B. megaterium PSA10 was 593,61 DUN/mg protein and PHB production from sago starch was 52,28 % of cell dry weight (CDW). FTIR analysis of the polymer indicated that the strain B.megaterium PSA10 was a potent PHB producer.

Somatic embryogenesis of Sandalwood (Santalum album L.) Toni Herawan; Mohammad Na'iem; Sapto Indrioko; Ari Indrianto
Indonesian Journal of Biotechnology Vol 19, No 2 (2014)
Publisher : Universitas Gadjah Mada

Sandalwood (Santalum album L.) is native species of Indonesia, especially in East Nusa Tenggara, is oneof the twenty two species of the genus Santalum in the world. Sandalwood is an important tree because it hashigh economic value can produce sandal oil these can be used for perfumes, cosmetics, pharmaceuticals, andare often used in religious ceremonies. In vitro particularly somatic embryogenesis has been widely appliedin the propagation of sandalwood. The Objective of this research is to obtain regeneration of sandalwoodthrough somatic embryogenesis using leaves explant from various clones. Medium for embryo induction is MS(Murashige and Skoog, 1962) solid medium containing treatment of 2,4-D (2,4-Dichlorophenoxyacetic acid)at various concentrations. To the media 0,15 mg /l kinetin, 40 g/l sucrose, and 2,5 g/l gelrite were added.Culture were incubated in the dark. Medium for Embryo development (maturation) is MS solid mediumcontaining treatment of BAP (Benzyl-amino-purine) at various concentrations. To the media 0,01 mg /l NAA(Napthalene-acetic-acid), 40 g/l sucrose, and 2,5 g/l gelrite were added. Culture were incubated in the light. Tostudy the specifi c structure of sandalwood somatic embryo early detection was conducted using histologicalanalysis. Results of anova showed that the clones, media, and interaction between clones with media did notsignifi cantly affect the development of sandalwood callus percentage. Results of anova showed that the clonesand BAP concentration signifi cantly effect to the embryo development of sandalwood.

Apoptosis and Phagocytosis Activity of Macrophages Infected by Mycobacterium tuberculosis Resistant and Sensitive Isoniazid Clinical Isolates Rachmawaty, Farida J.; Wibawa, Tri; Soesatyo, Marsetyawan H.N.E.
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (43.035 KB)

Mycobacterium tuberculosis (M.tb) is the main causative pathogen that cause the pulmonary tuberculosis.Intracellular M.tb was reported able to induce macrophages apoptosis, which may have crucial role in the regulationof immun response against M.tb infection. As an intracellular bacteria, M.tb able to live and replicate withinmacrophages. Phagocytosis is the first step to achieved this condition. The induction of macrophages apoptosis byINH resistant and sensitive M.tb clinical isolates, and H37Rv was studied. The macrophages apoptosis level weremeasured using an Ag-capture ELISA for histone and fragmented DNA (Cell Death Detection ELISAplus, RocheDiagnostic GmBH). Phagocytosis activity also analyzed, after staining using fluorescence dye (AcriFluorTM, ScientificDevice Lab.). The results showed that there was no significantly different between INH resistant and sensitive M.tbclinical isolates in respect their ability to induce apoptosis. The phagocytosis activity among the clinical isolates wasshown to be strain dependent, and undistinguishable between the Mtb clinical isolates. There was no associationbetween macrophages apoptosis level and the phagocytosis activity. These data suggested that among the virulentMtb clinical isolates, the ability to induce macrophages apoptosis and phagocytosis were consistently in comparablelevelKeywords: Mycobacterium tuberculosis, apoptosis, phagocytosis, macrophages, isoniazid

The Use of DNA Microsatellite Markers for Genetic Diversity Identification of Soybean (Glycine max (L) Meriil.) as a Supplementary Method in Reference Collections Management Nina Agusti Widaningsih; Edi Purwanto; N. Nandariyah; R. Reflinur
Indonesian Journal of Biotechnology Vol 19, No 2 (2014)
Publisher : Universitas Gadjah Mada

Large number of new soybean varieties are mostly derived from crosses of elite genotypes resulted ina narrowing of both the genetic diversity and the phylogenetic relationship between soybean varieties. Thus,discrimination among soybean varieties is becoming more diffi cult, especially when morphological traits wereapplied. In Plant Variety Protection (PVP) system, new varieties of soybeans including granted PVP right, localand breeding varieties registered in PVP offi ce were frequently increased, implicate on increasingly the numberof soybean varieties collections. To assist the management of varieties collections, a standard fi ngerprinting datais further needed. In comparison to the management of plant collection in the fi eld, molecular marker systemswhich are rapid, reliable, informative and relatively simple are continually sought for practical applications ingermplasm conservation, management and enhancement. This study aimed to identify the genetic diversity andphylogenetic relationship of soybean varieties that have earned PVP Right as well as local varieties and breedingvarieties registered in the PVP offi ce using microsatellite or simple sequence repeats (SSR) markers.This study was conducted in Molecular Biology laboratory, Indonesian Center for Agricultural Biotechnologyand Genetic Resources Research and Development (ICABIOGRAD) Bogor, from February to May 2013. The datawere analyzed using the genetic analysis package NTSYSpc 2.02i and PowerMarker V3.25. The result showed arelatively narrow genetic diversity among 45 varieties of soybean analyzed in present study which were indicatedby the small number of genotypes and total number of alleles (NA), and the low value of gene diversity and PICvalues (<0.75). Cluster analysis showed that the grouping varieties are not related to morphological characters butrelated to phylogeny relationship between varieties. Despite the group of varieties were not clustered in accordancewith morphological characteristics, SSR marker can be a powerful tool for discriminating varieties, so that it couldbe useful for initial varieties identity in conjunction with genetic diversity analysis.

Developmental Competence of Early Stage Porcine Embryos Cultured in Medium with Different Energy Substrate in vitro Ni Wayan Kurniani Karja; Kazuhiro Kikuchi; Mokhamad Fahrudin
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada

To elucidate the effect of energy requirement during the early embryonic development on their developmental ability to develop to blastocyst stage, in vitro fertilized (IVF) porcine one-cell embryos were cultured in modified North Carolina State University (NCSU)-37 supplemented with different energy substrate. Result indicated that the cleavage rate of embryos in Pyr-Lac and Gluc-Pyr-Lact groups was significantly higher than in those in Gluc group and Gluc-Rib group (P < 0.05). At Day 6 of culture, the highest proportion of embryos develop to the blastocyst stage was obtained in the presence of pyruvate-lactate only. In the medium with glucose, the addition of pyruvate-lactate or ribose slightly increased the proportion of embryos develop to the blastocyst stage, however the value were not significantly different form those obtained in the presence of glucose only. The mean cell number in blastocysts derived from Pyr-Lac and Gluc-Pyr-Lact groups were significantly higher than those in the Gluc group (P < 0.05). These results indicated that the presence of glucose only, as energy substrate, during the first 2 days of in vitro culture (IVC) caused a decrease in development of in vitro produced (IVP) porcine embryos to the blastocyst stage and mean cell number in blastocysts.

Polymorphism of Transcription Factor 7-Like 2 Gene and HOMA-β Level of Individuals With and Without Type 2 Diabetes Mellitus Family History Waode Astria Sahrani; Indwiani Astuti; Ahmad Hamim Sadewa
Indonesian Journal of Biotechnology Vol 19, No 2 (2014)
Publisher : Universitas Gadjah Mada

Family history has considered as a risk factor of type 2 diabetes. Transcription factor-7 like 2 (TCF7L2) hasrole to regulates insulin secretion and blood glucose homeostasis. The aim of current study was to determine thers7903146 polymorphism of TCF7L2 gene and homeostatic model assessment-β (HOMA-β) level on individual withand without type 2 Diabetes Mellitus (DM) family history. This work is a case-control study. Thirty six subjectswith type 2 DM family history and 36 subjects without type 2 DM family history were recruited. HOMA-βmeasure to analyze the insulin secretion. Polymorphisms of TCF7L2 gene was analyzed by using PCR-RFLPmethod. Statistical analysis was performed by using T-test, Mann-Whitney and Chi-square with signifi cancelevel 0.05. The frequency of the T allele of the cases were 4.2% and the controls were 2.8% (p=0.500). The oddratio was 0.649 (CI;95%:0.106-4.055). The HOMA-β levels of the cases were signifi cant low (132.56±62.48)compared with the controls (266.09±1.68) with p=0.000. The subjects with type 2 DM family history have asimilar frequency of having T alleles and CT/TT genotypes. The subjects with type 2 DM family history hassignifi cantly lower HOMA-β levels than subject without DM family history.

Molecular Identification of Phenol-Degrading and Biofi lm-Forming Bacteria from Wastewater and Peat Soil Arifah Khusnuryani; Erni Martani; Tri Wibawa; Jaka Widada
Indonesian Journal of Biotechnology Vol 19, No 2 (2014)
Publisher : Universitas Gadjah Mada

Phenol is hazardous aromatic pollutant which needs to be treated to reduce its hazardous effects.Bioremediation using bacteria which can form biofi lm offer an alternative wastewater treatment that is cheaperand environmentally safe. Eighteen strains of phenol-degrading and biofi lm-forming bacteria were isolatedfrom peat soil, also hospital and textile wastewater. Screening for phenol degradation ability of isolates wereperformed using Folin-ciocalteau reagent, while for biofi lm formation ability were performed using microtiterplate and crystal violet dye. Based on the ability to degrade phenol and to form biofi lm, four isolates (HP3,DOK135, DL120, andATA6) were choosen as phenol-degrading bacteria as well as biofi lm-forming bacteria.Based on phenotypic and genotypic characterization, isolate HP3 was highly similar to Rhodococcus equi strainDSM20307T, while DOK135 was highly similar to Enterobacter mori strain R18-2.The results also suggested thatDL120 and ATA6 could be classifi ed to the genus of Micrococcus and Bacillus respectively

The Susceptibility of katG Ser315Thr Mycobacterium tuberculosis mutant Against Anti Tuberculosis Drugs Ning Rintiswati; P. Praseno
Indonesian Journal of Biotechnology Vol 19, No 2 (2014)
Publisher : Universitas Gadjah Mada

Mycobacterium tuberculosis resistant strains is a matter of great concern for TB control Program since nocure for some multi drug resistance TB (MDR-TB) cases. These strains could spread around the world, makea great challenges for control measures. Mycobacterium tuberculosis strains resistance to anti tuberculosis drugsis mainly caused by the alteration in several genes encoding the molecular targets. Mutation of katG at codon315 especially Ser315Thr are responsible for INH resistance in a large proportion of TB cases. The aim of thisstudy is to know the frequency of of katG Ser315Thr of M. tuberculosis mutant and the pattern of resistance toAnti TB drugs. The study design was observational laboratoric. Eighty fi ve M. tuberculosis INH resistant clinicalisolates were screened for mutation of katGSer315Thr by using PCR-RFLP and DNA sequence analysis.Theresults showed that katG Ser 315Thr were identifi ed in 23 (27,05%) of INH resistance isolates. Frequency ofMDR among the katG Ser315Thr mutants almost double than the non mutant. The result suggested that thethe katG Ser315Thr mutation may play important role in the development of MDR-TB.

Page 15 of 52 | Total Record : 518

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