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ANNALES BOGORIENSES

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Published by Lembaga Ilmu Pengetahuan Indonesia

ISSN : 05178452 EISSN : 24077518 DOI : -

Core Subject : Health, Science, Agriculture, Engineering,

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Immunology & microbiology Medicine & Pharmacology

The Annales Bogorienses (ISSN: 0517-8452, E-ISSN: 2407-7518) is a peer-reviewed Journal that is published biannually. First published in 1955, it is now one of the oldest scientific journal in the nation. The Annales Bogorienses publishes original articles in basic and applied research as well as critical reviews and short communication in the fields of life sciences with the emphasis in biotechnology, molecular biology, and biochemistry.

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Articles 540 Documents

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Guide for Authors AB Vol 19 No 1 (2015) Dzikri, Muhamad
ANNALES BOGORIENSES Vol 19, No 1 (2015): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Construction and Expression of Immunotoxin Anti EGFRvIII scFv-HPR Conjugate in Pichia pastoris as A Targeted Drug Candidate for Cancer Therapy Yuliawati, Yuliawati; Soejoedono, Retno Damayanti; Fuad, Asrul Muhamad
ANNALES BOGORIENSES Vol 18, No 1 (2014): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/89

Epidermal growth factor receptor variant III (EGFRvIII) is a mutant of EGFR lacking 267 amino acids (exon-2 through 7) within its extracellular domain, resulting in the formation of a new epitope as a tumorspecific target. EGFRvIII is commonly found in GBM, breast, ovarian, prostate, and lung carcinomas. Antibodies or part of antibodies (e.g. single chain variable fragment or scFv) with specific activity against this receptor have been developed. These antibodies are internalized into the cell after receptor binding. Ribonucleases can be cytotoxic due to their inherent ability to degrade RNA, therefore causing cell death via inhibition of protein synthesis. The aim of this research is to construct, clone and express an immunotoxin-based conjugate combining an anti-EGFRvIII scFv with a HPRmut (human pancreatic ribonuclease mutant variant) in Pichia pastoris. The anti-EGFRvIII scFv gene was cloned into yeast expression vector pPICZÎ±A and fused with EGFP gene as a marker. The HPRmut gene was subsequently cloned at the C-terminal of the scFv::EGFP fusion, resulting in the scFv::EGFP::HPR fusion construct. The fusion construct was successfully obtained and nucleotide sequence of plasmid was verified. This construct was used to transform Pichia pastoris SMD 1168H. The gene fusion was successfully transformed and expressed in P. pastoris with a transformation efficiency of 102 cfu/Î¼g DNA. The transformed yeasts were screened on agar media containing up to 1000 Î¼g/ml zeocin. Yeast transformants showed green fluorescent due to the expression of EGFP gene. The recombinant proteins have been expressed and secreted from P. pastoris as showed by immunoblotting and SDS-PAGE analyses, then purified by affinity chromatography method. The selected yeast transformant produced at least 15.85 mg of purified protein per liter of yeast culture.

Propagation of Sukun (Artocarpus altilis (Parkinson) Fosberg) through In Vitro Shoot Proliferation Imelda, Maria; Wulansari, Aida; Sari, Laela
ANNALES BOGORIENSES Vol 13, No 1 (2009): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/28

Sukun (Artocarpus altilis (Parkinson) Fosberg) of the Moraceaae is a big tree which can grow to 15-20 m in height and native toÂ the Asia-Pasific region.Â Beside its delicious fruit, sukun is also known as aÂ traditional herbal medicinal plant in the region, including Indonesia.Â Nearly all parts of theÂ plant, such as roots, stems and leaves are believed by local communities to be capable of curing liver disease, hypertension, cardiac arrest, toothache, renal problem and even skin itchiness. The collaborative research between LIPI and PR China, on developing herbal medicines indicated that sukun has a great potential for treating cardiovascular disease.Â However, the availability of raw materials still poses a big constraint for the industry of herbal medicines. Generally, sukunÂ is propagated by root or stem cuttings, since in IndonesiaÂ sukun does not produce any seeds. However suchÂ method only produces limited planting materials. In general tissue culture propagated plants have many advantages, namely being clonal,Â Â free from pest and diseases, more uniform,Â andÂ allowing a high rate ofÂ plant multiplication. Therefore, the technique for sukun propagation has been developed by the LIPI Research Centre for Biotechnology. In this research the effects ofÂ 1-5 mg/l benzyl amino purine (BAP) and 20-40 mg/l adenine sulphate (AS) on shoot bud proliferation were investigated using lateral shoot buds on a modified Murashige and Skoog (MS) medium with addition of 150 ml/l coconut water(CW). Shoots were rooted on MS medium without plant growth regulators (PGRs). The results showed that the best medium forÂ in vitro shoot proliferation was a modified MS medium containing 2 mg/l BAP, 40 mg/l AS and 150 ml/l CW. The best medium for rooting is MS mediumÂ containing 1 mg indole butyric acid (IBA), producing roots within 3 weeks.Â Keywords : Sukun (Artocarpus altilis),Â in vitro shoot proliferation, Benzyl amino purine (BAP), Adenine sulphate (AS), coconut water (CW), Indole butyric acid (IBA)

Massive In Vitro Propagation Of Sandalwood Through Friable Embryogenic Callus Supatmi, Supatmi; Ardiyanti, Nurdiya; Rahman, Nurhamidar; Sudarmonowati, Enny
ANNALES BOGORIENSES Vol 20, No 1 (2016): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Sandalwood (Santalum album), which belongs to Santalaceae family, is a commercially important tree in Indonesia due to its many application. However,its population has significantly depleted since the planting materials using conventional methods are difficult to be provided. This study was conducted to mass propagate sandalwood using in vitro methods through friable embryogenic callus (FEC). The somatic embryos were formed using leaves cultured in MS +0.5 mg/l +1 mg/l indole acetic acid (IAA), and MS +1 mg/l IAA + 0.2 mg/l kinetin as well as 0.5 MS+1 mg/l Gibberellic acid (GA3). Primary somatic embryos (PSE) and secondary somatic embryos (SSE) then formed friable embryogenic callus when they were repetitively transferred to MS +1.5 mg/l BAP + 1.2 mg/l kinetin every 3 weeks. The maturation and regeneration of FEC was best done in the MS +1.5 mg/l BAP + 1.2 mg/l kinetin for 4-8 weeks. The acclimatization of sandalwood plantlets can be best conducted in the medium containing soil, sand and compos in ratio of 1:1:1 with the companion plant Murraya paniculata, which gave the best percentage of survival rate and the lowest percentage of falling leaves.

The Effect of Honey on Bacterial Growth, Protein Degradation ,Amino Acids Contents and Volatile Compounds of Milks at Storage Khusniati, Tatik; Widyastuti, Yantyati
ANNALES BOGORIENSES Vol 10, No 1 (2005): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/16

Pasteurized milks spoiled at refrigerated storage due to growth of psychrotrophic bacteria. Honey which contain antibacterial and aromatic compounds may be used as supplement to inhibit psychrotrophic bacteria activitiy . To know nutritional and flavor compounds of milks with and without honey, effect of honey on bacterial growth0protein degradation. amino acids and volatile compounds of stored milk were detected. Bacterial growth,protein degradation, amino acids and navor compounds were detected by total plate counts. formol titration,HPLC and GCMS, respectively. The results show that bacterial growth and protein degradations in honey milks were lower than that without honey. Bacterial growth (5.2x10^3 - 9,3x10^6 cfu/mL) and protein degradation (2,37-2.59%) in honey whole milks were lower than that (6.2x10^4 - 6.5x10^7 cfu/mL)(2.54-2.88%) in skim milks,respectively (P<0,05). At 10 days after use by date, changing between amino acids contents in whole milks withand without honey were more significant than that or skim milk (P<0.05);. and volatile compounds percentages in honey whole milks were higher than that without honey. while that in honey skim milks vice versa. Honey caused decreasing bacterial growth and protein degradation, changing aminoi acid contents and producing volatile compounds of stored milk, and honey whole milk were better than honey skim milks. Â

Overproduction and Purification of Soluble Recombinant Human Granulocyte Colony Stimulating Factor in Escherichia coli Using Thioredoxin as Fusion Agustiyanti, Dian Fitria; Retnoningrum, Debbie Sofie; Rachmawati, Heni; Fuad, Asrul Muhamad
ANNALES BOGORIENSES Vol 21, No 1 (2017): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ab.v21i1.294

Recombinant human Granulocyte Colony Stimulating Factor (G-CSF) has been produced in a soluble form in Escherichia coli BL21 (DE3) as a fusion protein. The open reading frame of G-CSF was synthetically constructed in previous work and was codon optimized for best expression in E. coli. In this research, the gene was fused to thioredoxin (Trx) at the N-terminal in pET32 vector. The purpose of this research was to optimize the overproduction and purification processes to obtain high yield recombinant protein in soluble form, and to characterize the Trx-G-CSF fusion protein. Overproduction was performed using IPTG induction method for 3 and 6 hours. The protein was purified by Ni-NTA affinity chromatography and separated using gradient concentration of imidazole. The purified protein was then characterized by SDS-PAGE and Western Blot analysis. Further, enterokinase was used to separate G-CSF from the fusion protein. The purified form of G-CSF was subsequently characterized using Western Blot and mass spectrometry using MALDI-TOF. The results showed that the fusion protein was successfully produced in soluble part as much as 48.25% were obtained after 3 hours of induction. The yield ofÂ fusion protein was 67.37% Â from total protein (229.65Â mg protein/L culture). The Western Blot analysis showed the G-CSF band at around 18.6 kDa. Mass spectrometry with MALDI-TOF/ TOF revealed that 25.86% of amino acid residue was recognized as part of human G-CSF sequence.Â

Improved Regeneration, Acclimatization and Shoot Cutting Production of âGebangâ Cassava Derived from Irradiated In Vitro Shoots Supatmi, Supatmi; Sudarmonowati, Enny
ANNALES BOGORIENSES Vol 16, No 2 (2012): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Gebang is an Indonesian local genotype which has been selected as superior genotype for low amylose Â cassava. Prior to induction of new mutants of this genotype, series of research have been conducted to improve regeneration and acclimatization as well as shoot cuttings production of irradiated in vitro shoots. Four dosage treatments of gamma ray irradiation i.e. those of at 0; 0.2; 1.0; 1.5; 2.0 krad were applied to 32 in vitro shoots multiplied from apical shoots of plants in the field. The highest multiplication rate and acclimatized shoots were obtained from shoots irradiated with 0.2 krad multiplied on various level of BAP added on MS medium and resulted that MS supplemented with 1 mg/L BAP was the best medium. The phenotypic variation was observed in shoots irradiated with 1.0; 1.5 and 2.0 krad while the ones irradiated with 0.2 krad performed normal appearances. Subsequent production of propagated young stem cuttings called âratooning systemâ decreased after the second cycle of propagation especially in the survival rate of the ones irradiated with 0.2 krad. The findings lead to the opportunity to produce cassava propagules derived from irradiated in vitro culture at a higher amount using Gebang genotype as a model.Keywords: cassava Gebang, irradiated shoot, young stem cutting/ratooning , acclimatization, in vitro.

Modification of Plasmids and Cry Genes of Bacillus Thuringiensis subsp. kurstaki Lid-I After Treatment With Ethymethanosulfonate (EMS) and UV Light Jusuf, Eddy
ANNALES BOGORIENSES Vol 9, No 2 (2004): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Bacillus thllringiensis subsp. kllrstaki HD-J is a potential insecticidal bacterium producing five type of a -endotoxin crystal proteins. This bacterium is widely commercialized due to its wide spectrum toxicity against both Lepidopteran and Dipteran larvae . The objective of this work was to create autolysin deficient mutant causing cell fails to lyse. This mutant yield intact cell s within spore and a -endotoxin crystal protein protected inside, from which an undamaged active bio-insecticide wou ld be obtained . Two methods of mutagenesis, 2ry( of ethylmethanosulfonate and 10, 25, and 50 second of UV light expo ure, resu lted in mor loss of motility) mutation. Observation showed that 14 of 20 survived mutants have lost some of its plasmids (varied from one to five). wh ile the other six maintained their plasmids. By employing the polymerase chain reaction (PCR) the change on the cry genes was studied. Â

Guide for Authors AB Vol 20 No 2 (2016) Dzikri, Muhamad
ANNALES BOGORIENSES Vol 20, No 2 (2016): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Optimizaton of Cationic Lipid Mediated Transfection of pEGFP-c1 and pJ-EPO Plasmids in Chinese Hamster Ovary (CHO) Cells Attached Culture for Transient and Stable Recombinant Human Erythropoietin (rhEPO) Expression Septisetyani, Endah Puji; Kusumawati, Arizah; Santoso, Adi
ANNALES BOGORIENSES Vol 19, No 1 (2015): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ab.v19i1.84

Cationic lipid is one of transfection agents which show high efficiency and low cytotoxicity. The transfection efficiencies may depend upon the type or amount of cationic lipids, the cell line or DNA plasmid being used for transfection. The purpose of this study was to find optimal condition for transfection of CHO-K1 and CHO-S cells with pJ-EPO plasmid (contain human epo gene) compared with pEGFP-c1 plasmid (contain gfp gene) by cationic lipid Lipofectamin 2000TM to generate stable transfectant expressing recombinant human erythropoietin (rhEPO). Optimization was carried out regarding the amount of lipofectamin, DNA concentration, and concentration of antibiotic Geneticin (G418) for selection of stable transfectants. Using standard amount of lipofectamin (10Âµl/well) in 6-well plate, highest expression level of green fluorescent protein (GFP) was shown after transfection of CHO-K1 cells with 4Âµg/well pEGFP-c1 while highest expression level of rhEPO was observed after transfection of CHO-K1 cells with 6, 8, and 10Âµg/well pJ-EPO plasmids. The data also indicated that optimal transfection conditions of CHO-K1 and CHO-S cells with pJ-EPO were shown with the use of 4Âµg/well DNA and 15Âµl lipofectamin, respectively. Concentration of G418 affected the expression where strongest rhEPO expression was shown at 1,000 ng/Âµl G418 concentration.Â

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