Indonesian Journal of Biotechnology
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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Cloning and Sequencing cDNA Encoding for Rhoptry-2 Toxoplasma Gondii Tachyzoite Local Isolate
M, Murwantoko;
Widada, Jaka;
Nuraini, Yani Lestari
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada
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Rhoptry protein belongs to an excretory and secretory antigens (ESAs) that play an important role during active penetration of parasite into the cell target. This protein an able Toxoplasma gondii to actively penetrate targeted cell, meanwhile ESAs protein stimulates intracellular vacuole modification. It is, therefore, after the parasite successfully enter the cell target then Granule (GRA) proteins are responsible for the formation of parasitophorus vacuole, which is protect the fusion with other intracellular compartments such as lysosomal vacuole. Consequently, this parasite is being able to survive and multiply at the cell target. The current study was aimed to clone and sequens cDNA encoding for ROP-2 of local isolated T. gondii tachizoite through DNA recombinant technique. Total ribonucleic acid (RNA) was isolated from tachyzoites of local isolated T. gondii that were grown up in Balb/c mice. Messenger RNA was isolated from total RNA using PolyAtract mRNA Isolation System. Messenger RNA was used as a template for synthesis cDNA using Riboclone cDNA Synthesis System AMV-RT. EcoRI adaptor from Riboclone EcoRI Adaptor Ligation System was added to Complementary DNA and than ligated to pUC19. Recombinant plasmid was transformed into E. coli (XL1-Blue). The transformed E. coli XL-1 Blue were plated on LB agar containing X-Gal, IPTG and ampicillin. Recombinant clones (white colony) were picked up and grown up in the LB medium at 37oC overnight. Expression of recombinant protein was analysed by immunoblotting in order to identify cDNA recombinant wich is express ESA of T. gondii local isolate. Recombinant plasmid were isolated using alkalilysis method and were elektroforated in 1% agarose gel. The isolated DNA recombinant plasmid was cut using Eco RI and then sequenced through Big Dye Terminator Mix AB1 377A Sequencer using M13 Forward and M13 Reverse primers. The conclusion of this results showed that the recombinant clone was coding for excretory and secretory protein which has molecular weight of 54 kDa. The DNA alignments of sequence from the cloned gene showed 97% homology with gene encoding for ROP-2 of T. gondii RH isolate., string),(99, en_US, subject, Toxoplasma gondii, tachizoite, ESA, complementary DNA, ROP2
PGV-1 is a Potent Antimitotic Agent
Widaryanti, Barinta;
Da’i, Muhammad;
Kawaichi, Masashi
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada
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Carcinogenesis may resulted from the malfunctioning of programmed cell death. Most of the anticancer drugs incurrent use induce apoptosis in susceptible cells. The fact that disparate agent interacting with different targets seemto induce cell death through some common mechanisms suggest that anticancer activity is determined by the abilityof inhibiting cell growth. Pentagamavunon-1 (PGV-1) is one of the curcumin analogues which showed to havepotency in inhibiting proliferation of T47D human breast carcinoma cells. The effects on T47D cells growth isassociated with cell cycle arrest in G2/M phase at the concentration of 2.5 ?M, followed by hyperploidy. The data onpolymerization assay, indicated that PGV-1 interact with tubulin in different manner from taxol. PGV-1 inhibittubulin polymerization on cell culture while taxol stabilized tubulin polymerization. Immunostainning data onPGV-1 treated cells showed slightly tubulin condensation, while taxol treated cells showed tubulin condensationdistinctly at 12 minutes after releasing from depolymerizing agent.In conclusion, PGV-1 represent a new microtubule inhibitor and has the potential to be developed for antimitoticdrugKey words: Pentagamavunon-1, T47D, tubulin
Actin Distribution in Lamina Neuralis During Cranial Neurulation of Wistar Rats Embryo (Rattus rattus)
Prahastuti, Indriayuni;
R, Issoegianti S.M.
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada
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such as craniorachisis and exencephaly. One of the processes is changing in lamina neuralis cells shape, which iscaused by actin microfilament rearrangement within lamina neuralis cells. To examine the distribution of actinmicrofilament during cranial neurulation Wistar rats embryo were used. Embryos were obtain at following days ofdevelopment; 8 days 18 hours, 9 days, 9 days 6 hours, 9 days 12 hours, 9 days 18 hours, and 9 days 20 hoursrespectively. Immunohistochemistry Avidin Biotin-peroxidase Complex (ABC) method was used to examine andidentify the distribution of actin in lamina neuralis cells. Light microscopic observation shows positive reaction foractin immunoreactivity in the apical surface of bending lamina neuralis cells. In contrast, actin is not observed in nonbendinglamina neuralis. Actin is not detected at 8 days 18 hours embryos. At 9 days embryos, positive reaction isobserved over the entire apical surface of lamina neuralis.Key words: Cranial neurulation, Actin, lamina neuralis, Rats embryo.
Cortisol and Estradiol Profile in Cross-bred Ettawa Does: The Effects of Body Condition Scoring (BCS).
Astuti, Puji;
S, Sunendar;
K, Suharto;
K, Asmarani;
A, Junaidi
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada
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Body Condition Scoring (BCS) is an estimation of the muscle and fat development of an animal. Thin ewes that are fighting to maintain their own body weight and low concentration of cortisol are not able to ovulate as ewes in a more desirable condition due to lack of oestradiol concentration. The aims of this research are to monitor the cortisol and oestradiol profile in Cross-bred ettawa does and to determine effect of BCS on the cortisol and oestradiol profile. Eight does were used in this research. These animals were devided equally into 2 groups based on Body Condition Scoring (BCS), namely BCS 2, which body weight range between 25-30 kgs as group I ( >n=4 ) and BCS 3 which consists of ettawa with body weight range between 33-40 kg as group II ( n=4 ). All animals were synchronized using implant of CIDR and PGF2alpha. Blood from jugular vein were collected every 3 and 6 hours as soon as oestrus until 72 hours. Serum contained cortisol and oestradiol then assayed using ELISA</div><div>method. Cortisol and oestradiol concentrations were compared between groups by T test. The results showed that average concentration of cortisol is 47.17 ± 42.19 ng/mL for BCS 2 and 112.40±74.41ng/mL for BCS 3 (P<0.05), whereas concentration of oestradiol is 72.25±30.62 pg/mL for BCS 2 and 145.72±100.18 pg/mL for BCS 3 (P<0.05). Either cortisol or oestradiol have very synchronized wave except 2 of animals from BCS 2 (50%), which has tendency to suppress each other. It was concluded that profile of cortisol and oestradiol hormone have a very similar pattern, and BCS can affect hormone profile.
Epitope Mapping of Fc gamma RIIa Monoclonal Antibodies
Sardjono, Caroline Tan;
Wines, Bruce;
Powel, Maree;
Hogarth, Mark
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada
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FcγRIIa (CD32) is an IgG receptor which has been shown to be important in autoimmune disease pathology. IV.3, 8.7, and 7.30 are anti-FcγRIIa monoclonal antibodies (mAbs), which block the interaction between FcγRIIa and complex IgG. In this study, the three mAbs were demonstrated to inhibit FcγRIIa function. The determination of the precise epitopes of the IV.3, 8.7, and 7.30 mAbs may become a potential approach for designing inhibitors for FcγRIIa. The epitope of IV.3, 8.7, and 7.30 were determined using chimeric receptors based on the extracellular domains of FcγRIIa and the FcεRI a chain. The epitopes for IV.3 was found to be mapped on amino acid residues 132-137, while 8.7 and 7.30 were on amino acid residues 112-119 and 157-162. Based on the crystal 3D model of FcγRIIa molecule, these amino acid sequences are clustered together forming a contiguous region within the ligand binding site of the receptor.
Identification of Thymocyte Subset by Multicolor Flow Cytometry ED LSR II FACSDriva ââ¬â FlowJo Software Analysis
H, Harapan;
W, Wienands;
I, Ichsan;
Indrayati, Ana
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada
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In their development, thymocytes express different cell surface molecules that important for identification of thymocyte subset. It’s not easy to detect this cell surface molecules to determine the thymocyte subpopulation for research. Here we used multicolor flow cytometry ED LSR II FACSDriva - FlowJo software to identify of thymocyte subset from thymocyte sample solution using several antibodies such as mouse anti rat CD2-FITC, mouse anti rat CD45RC-PE, mouse anti rat CD4-APC, mouse anti rat CD8á-PerCP, mouse anti rat CD3-Biotin + PE-Cy7 or APC-Cy7. We determined double negative and single positive thymocyt subset (CD4 or CD8), found that the double negative thymocyte subset express CD2 and CD45RC. It was useful to determine the thymocyte subset using multicolor flow sitometry ED LSR II FACSDriva - FlowJo software.
Actin Distribution in Lamina Neuralis During Cranial Neurulation of Wistar Rats Embryo (Rattus rattus)
Indriayuni Prahastuti;
S.M. Issoegianti R.
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada
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DOI: 10.22146/ijbiotech.7797
such as craniorachisis and exencephaly. One of the processes is changing in lamina neuralis cells shape, which iscaused by actin microfilament rearrangement within lamina neuralis cells. To examine the distribution of actinmicrofilament during cranial neurulation Wistar rats embryo were used. Embryos were obtain at following days ofdevelopment; 8 days 18 hours, 9 days, 9 days 6 hours, 9 days 12 hours, 9 days 18 hours, and 9 days 20 hoursrespectively. Immunohistochemistry Avidin Biotin-peroxidase Complex (ABC) method was used to examine andidentify the distribution of actin in lamina neuralis cells. Light microscopic observation shows positive reaction foractin immunoreactivity in the apical surface of bending lamina neuralis cells. In contrast, actin is not observed in nonbendinglamina neuralis. Actin is not detected at 8 days 18 hours embryos. At 9 days embryos, positive reaction isobserved over the entire apical surface of lamina neuralis.Key words: Cranial neurulation, Actin, lamina neuralis, Rats embryo.
Cortisol and Estradiol Profile in Cross-bred Ettawa Does: The Effects of Body Condition Scoring (BCS).
Puji Astuti;
D. Tri Widayati;
S. Sunendar;
K. Suharto;
Asmarani Kusumawati;
A. Junaidi
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada
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DOI: 10.22146/ijbiotech.7793
Body Condition Scoring (BCS) is an estimation of the muscle and fat development of an animal. Thin ewes that are fighting to maintain their own body weight and low concentration of cortisol are not able to ovulate as ewes in a more desirable condition due to lack of oestradiol concentration. The aims of this research are to monitor the cortisol and oestradiol profile in Cross-bred ettawa does and to determine effect of BCS on the cortisol and oestradiol profile. Eight does were used in this research. These animals were devided equally into 2 groups based on Body Condition Scoring (BCS), namely BCS 2, which body weight range between 25-30 kgs as group I ( >n=4 ) and BCS 3 which consists of ettawa with body weight range between 33-40 kg as group II ( n=4 ). All animals were synchronized using implant of CIDR and PGF2alpha. Blood from jugular vein were collected every 3 and 6 hours as soon as oestrus until 72 hours. Serum contained cortisol and oestradiol then assayed using ELISAmethod. Cortisol and oestradiol concentrations were compared between groups by T test. The results showed that average concentration of cortisol is 47.17 ± 42.19 ng/mL for BCS 2 and 112.40±74.41ng/mL for BCS 3 (P<0.05), whereas concentration of oestradiol is 72.25±30.62 pg/mL for BCS 2 and 145.72±100.18 pg/mL for BCS 3 (P<0.05). Either cortisol or oestradiol have very synchronized wave except 2 of animals from BCS 2 (50%), which has tendency to suppress each other. It was concluded that profile of cortisol and oestradiol hormone have a very similar pattern, and BCS can affect hormone profile.
Identification of Thymocyte Subset by Multicolor Flow Cytometry ED LSR II FACSDriva - FlowJo Software Analysis
H. Harapan;
W. Wienands;
I. Ichsan;
Ana Indrayati
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada
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DOI: 10.22146/ijbiotech.7794
In their development, thymocytes express different cell surface molecules that important for identification of thymocyte subset. It’s not easy to detect this cell surface molecules to determine the thymocyte subpopulation for research. Here we used multicolor flow cytometry ED LSR II FACSDriva - FlowJo software to identify of thymocyte subset from thymocyte sample solution using several antibodies such as mouse anti rat CD2-FITC, mouse anti rat CD45RC-PE, mouse anti rat CD4-APC, mouse anti rat CD8á-PerCP, mouse anti rat CD3-Biotin + PE-Cy7 or APC-Cy7. We determined double negative and single positive thymocyt subset (CD4 or CD8), found that the double negative thymocyte subset express CD2 and CD45RC. It was useful to determine the thymocyte subset using multicolor flow sitometry ED LSR II FACSDriva - FlowJo software.
Cloning and Sequencing cDNA Encoding for Rhoptry-2 Toxoplasma Gondii Tachyzoite Local Isolate
M. Murwantoko;
Jaka Widada;
Yani Lestari Nuraini
Indonesian Journal of Biotechnology Vol 13, No 1 (2008)
Publisher : Universitas Gadjah Mada
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DOI: 10.22146/ijbiotech.7795
Rhoptry protein belongs to an excretory and secretory antigens (ESAs) that play an important role during active penetration of parasite into the cell target. This protein an able Toxoplasma gondii to actively penetrate targeted cell, meanwhile ESAs protein stimulates intracellular vacuole modification. It is, therefore, after the parasite successfully enter the cell target then Granule (GRA) proteins are responsible for the formation of parasitophorus vacuole, which is protect the fusion with other intracellular compartments such as lysosomal vacuole. Consequently, this parasite is being able to survive and multiply at the cell target. The current study was aimed to clone and sequens cDNA encoding for ROP-2 of local isolated T. gondii tachizoite through DNA recombinant technique. Total ribonucleic acid (RNA) was isolated from tachyzoites of local isolated T. gondii that were grown up in Balb/c mice. Messenger RNA was isolated from total RNA using PolyAtract mRNA Isolation System. Messenger RNA was used as a template for synthesis cDNA using Riboclone cDNA Synthesis System AMV-RT. EcoRI adaptor from Riboclone EcoRI Adaptor Ligation System was added to Complementary DNA and than ligated to pUC19. Recombinant plasmid was transformed into E. coli (XL1-Blue). The transformed E. coli XL-1 Blue were plated on LB agar containing X-Gal, IPTG and ampicillin. Recombinant clones (white colony) were picked up and grown up in the LB medium at 37oC overnight. Expression of recombinant protein was analysed by immunoblotting in order to identify cDNA recombinant wich is express ESA of T. gondii local isolate. Recombinant plasmid were isolated using alkalilysis method and were elektroforated in 1% agarose gel. The isolated DNA recombinant plasmid was cut using Eco RI and then sequenced through Big Dye Terminator Mix AB1 377A Sequencer using M13 Forward and M13 Reverse primers. The conclusion of this results showed that the recombinant clone was coding for excretory and secretory protein which has molecular weight of 54 kDa. The DNA alignments of sequence from the cloned gene showed 97% homology with gene encoding for ROP-2 of T. gondii RH isolate.', 'string'),(99, 'en_US', 'subject', 'Toxoplasma gondii, tachizoite, ESA, complementary DNA, ROP2