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Widya Asmara
Bagian Mikrobiologi Fakultas Kedokteran Hewan UGM Yogyakarta
Veterinary

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ISOLATION, IDENTIFICATION AND ANALYSIS OF SHORT INTERSPERSED NUCLEAR ELEMENT (SINE) FROM VARANUS KOMODOENSIS Widya Asmara
Jurnal Sain Veteriner Vol 17, No 1 (1999)
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.8591

Abstract

All vertebrata genomes studied so far contain SINEs, that consti tute up to 5% of the total genome. Most of the SINE sequences studied so far are mammalian genomes. There Is very limited information about SINE sequences from rep­tiles, especially of Varanus family.Isolation of SINE sequences were carried out by con­structing the komodo genomic library in E. coli. Suspected positive clones were detected by genome to genome hybridiza­tion. The SINE sequences identification were carried out by enzymatic method, and the DNA sequences were analysed with the GENETYX-SV program-Up to now there are 2 SINE sequences from komodo genome have been isolated and identified. Their sequences homo logy is 85-2% From Database search analysis with the GenBank indicated that those SlNEs were not evolved from any spe­cific tRNA gene.
DETECTION PROTECTIVE ANTIGEN WITH MONOCLONAL ANTIBODY OF PASTEURELLA MULTOCIDA DOMPU ISOLATE Sri Murwani; Widya Asmara
Jurnal Sain Veteriner Vol 16, No 2 (1999)
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.8599

Abstract

Monoclonal antibodies can be used as specific probes for molecular structure such as drugs, hormones, receptor for hormones, or any other biologycally derived or biology-cally active materials. The potential usefulness of mono­clonal antibodies is in almost every field of biology and medicine.The study was carried out to produced monoclonal anti­bodies against outer membrane protein (OMP) Pasteurella multocida isolat Dompu, which can be used to isolate imunogenic OMP fractions.Three monoclonal antibodies which specifically react with OMP fraction of Pasteurella multocida isolat Dompu were obtained previously. Those monoclonal antibodies were pro­duced in Balb/c mouse. The antibodies production were measured by ELISA and were purified by affinity chromatograPhy.The monoclonal antibody were injected intraperitoneally into group of five rats. Two additional groups of five rats were injected with immune mouse serum and normal mouse serum, which served as a positive and negative controls respectively. All rats were challenged with 1 ml lethal dose of Pasteurella multocida 24 hours after the antibody injec­tion.The outer membrane protein was separated 12,5% SDS-PAGE. The region of gel containing the appropiate antigens (PI, P2, P3) were excised. Protein antigen was mixed with incomplete Freund's adjuvant (IFA) and injected intramuscu-lary into group of five rats at dose 0,2 ml per mouse. Two additional groups were injected with gel in IFA and OMP and IFA as a positive and negative controls. Each treated mouse and control was immunized four times, at weekly interval. Two weeks after final immunization rat were bled and their antibodies productions were measured with ELISA. All mice were then challenged with lethal dose of Pasteurella multo­cida.The result indicated that eventhough immungenic the PI, P2, P3 OMP fraction individually were not able to induce protective antibody. 
IMMUNOGENICITY AND PROTECTIVITY OF THE OUTER MEMBRANE PROTEIN BANDS OF PASTEURELLA MULTOCIDA ISOLATE U-6 Ignatius Mulyadi; Widya Asmara; B. Sardjono
Jurnal Sain Veteriner Vol 16, No 1 (1998)
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.8608

Abstract

The most immunogenic and protective fraction of outer membrane proteins of Pasieurella multacida has not well understood. This study was done to find our the immunogenicity and protectivity of outer membrane protein fractions of Pasteurella multicida isolat U-6 in mice. The organism was cultured in Luria Bertani broth. The outer membrane proteins were prepared according 10 Snipes et al., (1988), Electrophoresis of the protein was carried out in 10% of sodium dodecy 1-su I ph ale poly cry 1 amide gel electrophoresis, and stained with 0.3% Coomasie brilliant blue. The result indicated that there arc many protein bands with 5 major protein bands, those are having molecular weight of 20,1 kilo Dalton (kD), 27 kD, 36 kD, 52,7kD,and75 kD. Each protein bandof molecular weight of 27 kD, 36kD,and75kD was then cut, and homogenised, and was used as antigen to immunise the mice after added with adjuvant 1:1. Two weeks after immunisation all of the mice were bled and the sera were tested by ELISA. FourtyeigM hours from bleeding, ail of the immunised and control mice were challenged with 500 colony forming unit of virulent Pasteurefla mutticida isolate U-6, and observed their mortality. The result indicated thai the outer membrane protein bands of molecular weight of 36 kD and 75 kD were immunogenic (P>0,05), while the outer membrane protein band of 27 kD was less immunogenic (P>0.05), Band with moleculer weight of 27 kD produced 0% protection, bandof 36 kD produced 60% protection, and band of 75 kD produced 40% protection of mice challenged.
MONOCLONAL ANTIBODIES AGAINST OUTER MEMBRANE PROTEIN PASTEURELLA MULTOCIDA STRAIN DOMPU Sri Murwani; Widya Asmara
Jurnal Sain Veteriner Vol 16, No 1 (1998)
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.8618

Abstract

Monoclonal antibodies were produced against outer membrane protein of Pasteurella multocida strain Dompu. The bacterial isolate was obtained from [he bacteria! collection held at Microbiology Laboratory, Faculty of Veterinary Medicine, Gadjah Mada University. Irs purity and patogenicily has beeen tested.Killed PatTeurella multocida were used ro immunized 7 female, 8-10 week old of Balb/ c mice. Cell fusion between limphocytes and myeloma cells (2:1) were carried out by PEG stimulation and bybrids were cultured in  HAT medium.The OMP were prepared from sonicated organism using centrifugallon in the presense of sarcosyL These antigens were than used to select antibody producing hvbridomas ELISA and cloning of hybridomas were done by serial dilution method.Four hybridomas clones were obtained, till producing monoclonal antibodies which reacted specifically against OMP Pasietirelta multacida stain Dompu.
Co-Authors B. Sardjono Ignatius Mulyadi Sri Murwani Sri Murwani