Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate
Microneme 3 (MIC3) protein tachyzoites Toxoplasma gondii is one of protein which plays an important role<br />during cell host invasion. Gene encoding MIC3 protein has been studied and it was suggested a potent vaccine<br />candidate against Toxoplasma gondii infection. The aim of this r...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Universitas Gadjah Mada, Yogyakarta
2015-10-01
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| Series: | Indonesian Journal of Biotechnology |
| Online Access: | http://journal.ugm.ac.id/ijbiotech/article/view/7410 |
| _version_ | 1811292683768954880 |
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| author | Wayan T. Artama1 Ni Nyoman Ayu Dewi Didik Tulus Subekti |
| author_facet | Wayan T. Artama1 Ni Nyoman Ayu Dewi Didik Tulus Subekti |
| author_sort | Wayan T. Artama1 |
| collection | DOAJ |
| description | Microneme 3 (MIC3) protein tachyzoites Toxoplasma gondii is one of protein which plays an important role<br />during cell host invasion. Gene encoding MIC3 protein has been studied and it was suggested a potent vaccine<br />candidate against Toxoplasma gondii infection. The aim of this research is to clone and sequence the gene encoding<br />MIC3 protein of tachyzoites Toxoplasma gondii local isolate by amplification using polymerase chain reaction with<br />specific primers. The amplified DNA fragment was cloned into pGEM-T and transformed into E. coli XL-1 Blue by<br />heat shock method. Recombinant plasmids were isolated using alkali lysis method and analyzed by digestion<br />using restriction endonuclease enzymes PstI, HindIII, NcoI and EcoRV. The recombinant plasmids then sequenced<br />to find out the nucleotide sequence of insert gene by ABIPRISM 377 DNA Sequencer. The DNA sequence then<br />were analyzed by computer software for alignment. The result showed that transformation in E. coli XL-1 Blue by<br />pGEM-T produced one clone that was encoding MIC3 protein. Analysis of 489 bp from 5’ and 447 from 3’ of gene<br />sequence showed 97-98% homology with gene encoding for MIC3 protein of RH isolate.<br />Keywords: MIC3 protein, Toxoplasma gondii, tachyzoite, recombinant DNA |
| first_indexed | 2024-04-13T04:49:04Z |
| format | Article |
| id | doaj.art-b6495f0088984e6b9577613348b01271 |
| institution | Directory Open Access Journal |
| issn | 0853-8654 2089-2241 |
| language | English |
| last_indexed | 2024-04-13T04:49:04Z |
| publishDate | 2015-10-01 |
| publisher | Universitas Gadjah Mada, Yogyakarta |
| record_format | Article |
| series | Indonesian Journal of Biotechnology |
| spelling | doaj.art-b6495f0088984e6b9577613348b012712022-12-22T03:01:44ZengUniversitas Gadjah Mada, YogyakartaIndonesian Journal of Biotechnology0853-86542089-22412015-10-011016243Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local IsolateWayan T. Artama1Ni Nyoman Ayu DewiDidik Tulus SubektiMicroneme 3 (MIC3) protein tachyzoites Toxoplasma gondii is one of protein which plays an important role<br />during cell host invasion. Gene encoding MIC3 protein has been studied and it was suggested a potent vaccine<br />candidate against Toxoplasma gondii infection. The aim of this research is to clone and sequence the gene encoding<br />MIC3 protein of tachyzoites Toxoplasma gondii local isolate by amplification using polymerase chain reaction with<br />specific primers. The amplified DNA fragment was cloned into pGEM-T and transformed into E. coli XL-1 Blue by<br />heat shock method. Recombinant plasmids were isolated using alkali lysis method and analyzed by digestion<br />using restriction endonuclease enzymes PstI, HindIII, NcoI and EcoRV. The recombinant plasmids then sequenced<br />to find out the nucleotide sequence of insert gene by ABIPRISM 377 DNA Sequencer. The DNA sequence then<br />were analyzed by computer software for alignment. The result showed that transformation in E. coli XL-1 Blue by<br />pGEM-T produced one clone that was encoding MIC3 protein. Analysis of 489 bp from 5’ and 447 from 3’ of gene<br />sequence showed 97-98% homology with gene encoding for MIC3 protein of RH isolate.<br />Keywords: MIC3 protein, Toxoplasma gondii, tachyzoite, recombinant DNAhttp://journal.ugm.ac.id/ijbiotech/article/view/7410 |
| spellingShingle | Wayan T. Artama1 Ni Nyoman Ayu Dewi Didik Tulus Subekti Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate Indonesian Journal of Biotechnology |
| title | Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate |
| title_full | Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate |
| title_fullStr | Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate |
| title_full_unstemmed | Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate |
| title_short | Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate |
| title_sort | cloning gene encoding micronema 3 mic3 protein of tachyzoite toxoplasma gondii local isolate |
| url | http://journal.ugm.ac.id/ijbiotech/article/view/7410 |
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