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Effect of maturation periods and leukaemia inhibitory factor on in vitro bovine embryo development Margawati, Endang Tri
Indonesian Journal of Animal and Veterinary Sciences Vol 1, No 3 (1995)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (612.166 KB) | DOI: 10.14334/jitv.v1i3.26

Abstract

The period of in vitro maturation,~20 vs 24 hours) with of without supplementation of :,~uk_ni;ia inhib toryy factor (LIF) (0, 500, 1000 or 2000 U/ml) was studied on bovine embryo development in vitro in a 2 x 4 factotial experiment w id oestgnoo i, a randomized block design . A total of 870 bovine oocytes were used . Besides embryo development, cell numbers of blastocysts v. : ;re also co , mted in order to study the quality of the embryos . Oocytes were matured in a modified TCM199 medium containing 10 ug/nil of FS-14 and LP., I 1~ t1L rstradiol, fertilized in TALP and cultured in SOF/AABSA medium. There was no interaction between maturation periods and LIF doses on embryo development (P>0 .05). Maturation periods, however, affected (P<0.05) blastocyst rates but did not for cleavage o, oocytes and the percentage of oocytes that developed into blastocysts . LIF doses during in vitro maturation did not affect embryo development (P>0 .05) . Cell numbers of blastocysts were also not affected by maturation periods and LIF doses (P<0 .05), however 20 h in vitro maturation and supplementation of LIF doses tended to increase the cell numbers. This study suggests that 20 h maturation increases blastocyst rates and that supplementation with LIF during maturation does not affect the quality of embryos produced in vitro. Key words : in vitro fertilization, oocyte, cleavage, blastocyst, cell numbers of blastocyst
IDENTIFIKASI VIRUS PENYAKIT JEMBRANA PADA SAPI BALI MENGGUNAKAN PENANDA MOLEKULER GEN env SU [Identification of Jembrana Disease Virus by Using a Molecular Marker of env SU Gene in Bali Cattle] Indriawati, Indriawati; Margawati, Endang Tri; Ridwan, Muhammad
BERITA BIOLOGI Vol 12, No 2 (2013)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (182.055 KB) | DOI: 10.14203/beritabiologi.v12i2.534

Abstract

Up to present, detection of Jembrana disease virus has been identified through serological test. Advances in molecular biology has enabled to detect Jembrana disease virus earlier, quicker and more accurate by application of molecular markers.The aim of this study was to identify Jembrana disease by using molecular marker of env SU gene in Bali cattle.Total RNA of Jembrana disease virus (7732bp) was collected from spleen of Bali cattle suspected Jembrana disease by using RNEasy Protect Mini Kit (QIAGEN). A pair of specific primers was designed from Jembrana viral genome (env SU) that accessed through a GenBank with Accession Number of U21603.A kit of Access Quick RT-PCR System (PROMEGA) was used for Reverse-Transcriptase-PCR (RT-PCR). The RT-PCR products were visualized on 2% agarose gel.The result showed a single band with the size of ± 900bp in all samples. This size indicated that env SU gene was existed in the examined spleen samples. This finding suggests that a molecular marker could be used accurately to identify the env SU gene in JDV of Bali cattle.
Enhancing Local Tropical Beef Cattle Production Supporting Food Security in Indonesia Margawati, Endang Tri
Proceeding INTERNATIONAL SEMINAR IMPROVING TROPICAL ANIMAL PRODUCTION FOR FOOD SECURITY PROCEEDING INTERNATIONAL SEMINAR
Publisher : Proceeding INTERNATIONAL SEMINAR IMPROVING TROPICAL ANIMAL PRODUCTION FOR FOOD SECURITY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (684.572 KB)

Abstract

Enhancing Local Tropical Beef Cattle Production Supporting Food Security inIndonesia
Effect of Ethanol and IPTG on the Recombinant Jembrana Trans-Activator of Transcriptation Protein Expression Indriawati, Indriawati; Salfia, Mega; Susanti, R; Margawati, Endang Tri
Biosaintifika: Journal of Biology & Biology Education Vol 10, No 3 (2018): December 2018
Publisher : Department of Biology, Faculty of Mathematics and Sciences, Semarang State University . Ro

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (526.116 KB) | DOI: 10.15294/biosaintifika.v10i3.15596

Abstract

Jembrana diseases are caused by Jembrana Diseases Virus (JDV). The previous study showed that Jembrana Trans-Activator of Trancriptation (JTAT) recombinant protein is effective as a vaccine for Jembrana diseases. The production of JTAT protein needs to be optimized to obtain a higher amount of vaccine. High expression of JTAT protein will produce a high vaccine product. This study aimed to examine the effect of the addition of ethanol and IPTG in E. coli media on the expression of JTAT recombinant protein. This research was experimental research with factorial RAL design with a variation factor of ethanol and IPTG. Qualitatively, the induction of each IPTG, ethanol and interaction between the two could induce the expression of JTAT protein and could be identified with SDS-PAGE at ±11.8 kDa. Statistically, the induction of IPTG, ethanol and interaction between the two were not significantly different. Qualitative and quantitative data show that ethanol can induce JTAT protein expression. This result can be used as a preliminary study to test the effectiveness of ethanol as a substitute for IPTG in inducing the recombinant protein expression.
Producing the Greatest Good for Greatest Numbers-Implementation of Utilitarianism Principle: The Case Study of Producing Recombinant Protein of JDV Margawati, Endang Tri
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 15, No 2 (2010): June 2010
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (375.236 KB) | DOI: 10.24002/biota.v15i2.2729

Abstract

Advanced technology in molecular biology often uses microorganism, consequently, researcher should have a responsibility in producing of laboratory products safely both for human and their environment. This presentation was intended (1) to report recombinant protein research in the Jembrana Disease Virus (JDV); (2) to identify relevancy of the ethics towards the research of recombinant protein and (3) to discuss relationship of utilitarianism principle with the development of the recombinant protein. The Jembrana disease is an infectious virus caused by a virus classified as retrovirus of Retrovidae family. The disease only attacks Bali cattle (Bos javanicus) that caused about 20% mortality rate. Up to present, crude vaccine from lymph organ of acute infected Bali cattle is often used for vaccination. Development of the Jembrana vaccine was attempted to increase the availability of qualified Jembrana vaccine by recombinant DNA approaches subsequently could be used as vaccine substances. This article was presented with much bioethics issues in associated with recombinant protein research and other examples of related research which use micro-organism in their investigation. It is expected that bioethics could be a restrain for researchers who deal with advanced technology in their investigation.
Expression and Characterization of Recombinant Protein of J-SU pGEX either by Single or Double Cell Lysis Margawati, Endang Tri; Ridwan, Muhamad
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 14, No 3 (2009): October 2009
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (201.705 KB) | DOI: 10.24002/biota.v14i3.2579

Abstract

Penelitian ini dimaksudkan untuk optimasi produk protein rekombinan Superficial Unit dari virus Jembrana (JSU) yang dieksperikan melalui pemecahan sel secara tunggal dan ganda dengan sistem pGEX dalam skala flask 100ml media kultur. Dua metode pemecahan sel yang digunakan yaitu Freeze and Thaw (FT) sebagai pemecahan tunggal dan gabungan FT dan Sonikasi sebagai pemecahan ganda. Sel inang (E. Coli pembawa konstruk JSU pGEX) ditumbuhkan dengan induksi IPTG pada 37oC dengan pengocok berkecepatan 200rpm sampai mencapai kepadatan sel 0,8. Sel atau pelet dikoleksi dengan sentrifugasi, pelet dipecah dengan 2 perlakuan pemecahan sel tunggal dan ganda. Hasil pemecahan sel disentrifugasi untuk dikoleksi peletnya sebagai inclusion body. Solubilisasi dilakukan terhadap inclusion body dengan solubilisasi buffer dan diperoleh substrat protein JSU kemudian dimurnikan melalui Gluthation sepharose 4B (500μl resin) dengan metode batch capture. Hasil karakterisasi dengan SDS PAGE dan Western Blotting menunjukkan ukuran protein JSU pGEX yang tepat yaitu 60kDa pada kedua sistem pemecahan sel. Namun demikian, pemecahan sel secara tunggal menghasilkan protein murni JSU pGEX lebih besar (0.812ng/ul) dibanding pemecahan sel secara ganda (0.486ng/ul). Dari penelitian ini dapat disimpulkan bahwa protein rekombinan JSU pGEX terekspresi lebih baik dengan metode pemecahan sel Freeze and Thaw.
PENGUJIAN PENCEMARAN DAGING BABI PADA BEBERAPA PRODUK BAKSO DENGAN TEKNOLOGI PCR: PENCARIAN SISTEM PENGUJIAN EFEKTIF Margawati, Endang Tri; Ridwan, Muhamad
BERITA BIOLOGI Vol 10, No 1 (2010)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v10i1.2055

Abstract

Entering globalization market, Indonesian government could not reject any import of food products from overseas. To anticipate the possibility of porcine contaminants into processed meat products of imported food such as meat or chicken ball, sausage, tin meat etc., it is important to apply laboratory research on such particular matter in regard to ethical and certain religious concern. This study was intended to identify the possibility of porcine contaminants into either processed meat products or fresh meat.A technique of polymerase chain reaction (PCR) was applied and PCR optimizing was conducted in advanced to obtain the right annealing temperature.Positive control of fresh pork meat was amplified to get porcine Leptin size which is 152bp fragment. Five samples of 4 meat balls and one fresh beef meat were individually collected for their DNA by either from minced or mashed after liquid nitrogen exposure then followed with a series of DNA extraction steps. PCR was assigned by using a specific primer of Leptin gene for porcine identification.Visualization of Leptin fragment was applied either on 1%, 2% of agarose gel or 10-20% gradient acrylamide gel.The result showed that all sample applied were not identified for containing porcine contaminants while positive control was on the right size of 152bp of Leptin gene. Specific primer used in this study was proved that there was not identified porcine Leptin gene on the negative control (fresh beef meat). This study suggests that a method of PCR is a simple analytical method for identification of porcine contaminants and visualization on 2% agarose gel is a cheaper and quicker method while by gradient acrylamide gel showing more clear band however this method is time consuming and expensive.
In Vitro Development of Ovine Embryos Following Maturation Under Limited CO2 ENDANG TRI MARGAWATI
HAYATI Journal of Biosciences Vol. 12 No. 3 (2005): September 2005
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (58.347 KB) | DOI: 10.4308/hjb.12.3.112

Abstract

An experiment was conducted to examine the influence of CO2 during in vitro oocyte maturation on the in vitro ovine embryo development. Three treatments of CO2 were subjected to the oocyte development. Those were 2h gasses prior to maturation in incubator (T1); without CO2 either prior to or over maturation (T2) and CO2 exposure both prior to and over 22h maturation (T3). A total of 324 oocytes were used. Putative zygotes were cultured for seven days and evaluated for their developmental stage. Presence of CO2 (T3) increased the proportion of oocytes reaching Metaphase II ( 66.50 + 3.5%; p
Quantitative Trait Loci (QTL) Analysis for Production Traits of Birth Weight and Weight 360 days in Backcross Sheep ENDANG TRI MARGAWATI; HERMAN WILLEM RAADSMA; HARIMURTI MARTOJO; SUBANDRIYO SUBANDRIYO; MULADNO MULADNO
HAYATI Journal of Biosciences Vol. 13 No. 1 (2006): March 2006
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (38.729 KB) | DOI: 10.4308/hjb.13.1.31

Abstract

Four half-sib families (n = 382) consisting predominantly of ITT x Merino x Merino backcross progeny, including some F2 progeny were used to analyze QTL for two production traits (Birth weight = BW1 and Body weight at 360 days = BW360). The study exploited differences in weight performance between the Merino and ITT sheep. A total of 141 informative microsatellite markers were used in a genome-wide scan covering the 26 autosomal sheep chromosomes. QTL analysis was conducted online using QTL Express. This study reports the effect of QTL for birth weight on Chromosomes 5 (p < 0.05) at 112cM (0cM-128cM). Location of candidate genes for birth weight was predicted at the region of flanking markers MCM527-BMS1247. A QTL for BW360 days existed on Chromosome 18 (p < 0.01) at 104cM (25.0-125cM). Location of candidate genes related to production traits for body weight 360 days was predicted at the segment of flanking markers of CSSM018-TMR1. Only the QTL on Chromosome 18 retained significance (p < 0.01) under experiment-wide significance testing. This QTL region is being examined for candidate genes by investigating to the homologous human chromosomal segments. Key words: Quantitative trait loci, production traits, birth weight, weigh 360, backcross sheep
Maturing Singly Versus in Groups on Ovine Oocyte Meiosis Endang Tri Margawati
Media Veteriner Vol. 5 No. 3 (1998): Media Veteriner
Publisher : Media Veteriner

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (525.528 KB)

Abstract

The effect of maturing systems (single vs groups) of ovine oocytes was studied to examine the oocyte development in vitro. Oocytes were aspirated from follicle ovaries collected at local abattoirs Cibinong West Java, using a 18-G needle. Aspiration medium consisted of H 199 + 2 % FCS + 50 μg/ml Heparin and the maturating medium (IVM) consisted of B 199 - 10 % FCS + 10 μg/ml FSH + 10 μg/ml hCG + 1 μg/ml Estradiol. The oocytes collected were divided into three groups and treated separately as follows: T1) oocytes were matured singly in 50 μl IVM medium. T2) every five oocytes was matured in 50 μl IVM medium and T3) every 10 oocytes was matured in 50 μl IVM medium. All oocytes were maintained in incubator with 5 % CO, and high humidity at 38 °C for 20 h. The resulting ova were stained in 1 % lacmoid then examined for meiosis division under a microscope. There was a signiticant effect among treatments on the proportion oocytes reaching metaphase II (P