AMPLIFIKASI, SUBKLONING DAN SEKUENSING GEN PENYANDI PROTEIN INTERFERON GAMMA (IFN-γ) UNTUK PENGEMBANGAN DIAGNOSIS TUBERKULOSIS PADA ANJING
Tuberculosis is a disease caused by close related Mycobacterium strains such as M.tuberculosis, M.bovis, M.africanum, and M.microti which are known as M.tuberculosis complex (MTC). Tuberculosis (TB) is mainly described by the presence of tubercle or granulomatous inflammation which could happen in h...
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[Yogyakarta] : Universitas Gadjah Mada
2012
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id-ugm-repo.980642016-03-04T08:47:46Z https://repository.ugm.ac.id/98064/ AMPLIFIKASI, SUBKLONING DAN SEKUENSING GEN PENYANDI PROTEIN INTERFERON GAMMA (IFN-γ) UNTUK PENGEMBANGAN DIAGNOSIS TUBERKULOSIS PADA ANJING , Mei Ita Riyanti , Prof. Dr. drh. Ida Tjahajati, M.P. ETD Tuberculosis is a disease caused by close related Mycobacterium strains such as M.tuberculosis, M.bovis, M.africanum, and M.microti which are known as M.tuberculosis complex (MTC). Tuberculosis (TB) is mainly described by the presence of tubercle or granulomatous inflammation which could happen in human as well as many species of animals. Tuberculosis in small animal gains more attention because it can endanger human health. This research aims to develop a new diagnostic tool in identification of tuberculosis infection in dogs by subcloning of interferon gamma protein coding gene. Obtained interferon gamma protein can be used to produce primary and secondary antibodies which can be used in tuberculosis diagnosis in dogs by ELISA method. The research started by amplification of IFN-γ from mRNA obtained from Tjahajati�s research (2008) to DNA by RT-PCR. Obtained DNA were ligated to plasmid vector pET SUMO and transformed to One Shot® Chemically Competent E. coli Mach 1 (One Shot® Mach1�-T1). Growing colonies were recultured to liquid Luria Bertani (LB) media containing kanamycin. The next step is isolating plasmid using High Pure Plasmid Isolation Kit (Roche). Isolated plasmids were tested using PCR with canine IFN-γ primer. Product of PCR than being sequenced and analysed with Mega version 4.0. The results showed that bacteria colonies successfully grown on LB plat media and grown in re-cultured liquid LB media. After plasmid isolation and PCR testing, positive results were marked by 541 bp band in electrophoresis equal to IFN-γ gene width. The sequence�s result show 91% similarity between gene canine IFN-γ from cloning and gene canine IFN-γ from Gene bank. [Yogyakarta] : Universitas Gadjah Mada 2012 Thesis NonPeerReviewed , Mei Ita Riyanti and , Prof. Dr. drh. Ida Tjahajati, M.P. (2012) AMPLIFIKASI, SUBKLONING DAN SEKUENSING GEN PENYANDI PROTEIN INTERFERON GAMMA (IFN-γ) UNTUK PENGEMBANGAN DIAGNOSIS TUBERKULOSIS PADA ANJING. UNSPECIFIED thesis, UNSPECIFIED. http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=54352 |
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ETD , Mei Ita Riyanti , Prof. Dr. drh. Ida Tjahajati, M.P. AMPLIFIKASI, SUBKLONING DAN SEKUENSING GEN PENYANDI PROTEIN INTERFERON GAMMA (IFN-γ) UNTUK PENGEMBANGAN DIAGNOSIS TUBERKULOSIS PADA ANJING |
description |
Tuberculosis is a disease caused by close related Mycobacterium strains
such as M.tuberculosis, M.bovis, M.africanum, and M.microti which are known as
M.tuberculosis complex (MTC). Tuberculosis (TB) is mainly described by the
presence of tubercle or granulomatous inflammation which could happen in
human as well as many species of animals. Tuberculosis in small animal gains
more attention because it can endanger human health. This research aims to
develop a new diagnostic tool in identification of tuberculosis infection in dogs by
subcloning of interferon gamma protein coding gene. Obtained interferon gamma
protein can be used to produce primary and secondary antibodies which can be
used in tuberculosis diagnosis in dogs by ELISA method.
The research started by amplification of IFN-γ from mRNA obtained from
Tjahajati�s research (2008) to DNA by RT-PCR. Obtained DNA were ligated to
plasmid vector pET SUMO and transformed to One Shot® Chemically
Competent E. coli Mach 1 (One Shot® Mach1�-T1). Growing colonies were recultured
to liquid Luria Bertani (LB) media containing kanamycin. The next step
is isolating plasmid using High Pure Plasmid Isolation Kit (Roche). Isolated
plasmids were tested using PCR with canine IFN-γ primer. Product of PCR than
being sequenced and analysed with Mega version 4.0.
The results showed that bacteria colonies successfully grown on LB plat
media and grown in re-cultured liquid LB media. After plasmid isolation and PCR
testing, positive results were marked by 541 bp band in electrophoresis equal to
IFN-γ gene width. The sequence�s result show 91% similarity between gene
canine IFN-γ from cloning and gene canine IFN-γ from Gene bank. |
format |
Theses and Dissertations NonPeerReviewed |
author |
, Mei Ita Riyanti , Prof. Dr. drh. Ida Tjahajati, M.P. |
author_facet |
, Mei Ita Riyanti , Prof. Dr. drh. Ida Tjahajati, M.P. |
author_sort |
, Mei Ita Riyanti |
title |
AMPLIFIKASI, SUBKLONING DAN SEKUENSING GEN PENYANDI
PROTEIN INTERFERON GAMMA (IFN-γ) UNTUK PENGEMBANGAN
DIAGNOSIS TUBERKULOSIS PADA ANJING |
title_short |
AMPLIFIKASI, SUBKLONING DAN SEKUENSING GEN PENYANDI
PROTEIN INTERFERON GAMMA (IFN-γ) UNTUK PENGEMBANGAN
DIAGNOSIS TUBERKULOSIS PADA ANJING |
title_full |
AMPLIFIKASI, SUBKLONING DAN SEKUENSING GEN PENYANDI
PROTEIN INTERFERON GAMMA (IFN-γ) UNTUK PENGEMBANGAN
DIAGNOSIS TUBERKULOSIS PADA ANJING |
title_fullStr |
AMPLIFIKASI, SUBKLONING DAN SEKUENSING GEN PENYANDI
PROTEIN INTERFERON GAMMA (IFN-γ) UNTUK PENGEMBANGAN
DIAGNOSIS TUBERKULOSIS PADA ANJING |
title_full_unstemmed |
AMPLIFIKASI, SUBKLONING DAN SEKUENSING GEN PENYANDI
PROTEIN INTERFERON GAMMA (IFN-γ) UNTUK PENGEMBANGAN
DIAGNOSIS TUBERKULOSIS PADA ANJING |
title_sort |
amplifikasi, subkloning dan sekuensing gen penyandi
protein interferon gamma (ifn-î³) untuk pengembangan
diagnosis tuberkulosis pada anjing |
publisher |
[Yogyakarta] : Universitas Gadjah Mada |
publishDate |
2012 |
url |
https://repository.ugm.ac.id/98064/ http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=54352 |
_version_ |
1681230287575449600 |