Construction of a plant transformation vector containing a partial isochorismate syhthase cDNA
Isochorismate synthase (ICS) is an enzyme that catalyzes chorismate into isochorismate. Isochorismate is a channel for the production of secondary metabolites such as anthraquinone and phylloquinones. A 1502 bp fragment partial ICS cDNA was amplified from pGEM-T vector through PCR using a set of pri...
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| Format: | Final Year Project Report / IMRAD |
| Language: | English English |
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Universiti Malaysia Sarawak, (UNIMAS)
2012
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| Online Access: | http://ir.unimas.my/id/eprint/8125/ http://ir.unimas.my/id/eprint/8125/1/Construction%20of%20A%20Plant%20Transformation%20Vector%20Containing%20A%20Partial%20Isochorismate%20Synthase%20cDNA%20%2824pgs%29.pdf http://ir.unimas.my/id/eprint/8125/8/Tai%20Ker%20Shing.pdf |
| _version_ | 1848836306183913472 |
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| author | Tai, Ker Shing |
| author_facet | Tai, Ker Shing |
| author_sort | Tai, Ker Shing |
| building | UNIMAS Institutional Repository |
| collection | Online Access |
| description | Isochorismate synthase (ICS) is an enzyme that catalyzes chorismate into isochorismate. Isochorismate is a channel for the production of secondary metabolites such as anthraquinone and phylloquinones. A 1502 bp fragment partial ICS cDNA was amplified from pGEM-T vector through PCR using a set of primers. A binary vector, pGSA 1l31, was isolated from E. coli XLI Blue cells and was linearized using BamH 1 restriction endonuclease at the BamH 1 restriction site. Both cDNA and pGSA 1131 plasmid were subjected to end-blunting procedure. The partial ICS cDNA was ligated into pGSA 1131 binary vector through blunt end ligation to form a recombinant vector. Ligation mixture was transformed into competent E. coli XLI Blue cells via heat shock method. Single colonies were grown from a LB agar plate and five colonies were chosen and were heat disrupted to release their plasmid content. Colony PCR technique was performed using primers designed from conserved ICS domain. An amplified 800 bp fragment of the internal confirms the presence of the partial ICS cDNA in the colonies. |
| first_indexed | 2025-11-15T06:21:40Z |
| format | Final Year Project Report / IMRAD |
| id | unimas-8125 |
| institution | Universiti Malaysia Sarawak |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T06:21:40Z |
| publishDate | 2012 |
| publisher | Universiti Malaysia Sarawak, (UNIMAS) |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | unimas-81252024-04-30T04:49:51Z http://ir.unimas.my/id/eprint/8125/ Construction of a plant transformation vector containing a partial isochorismate syhthase cDNA Tai, Ker Shing Q Science (General) Isochorismate synthase (ICS) is an enzyme that catalyzes chorismate into isochorismate. Isochorismate is a channel for the production of secondary metabolites such as anthraquinone and phylloquinones. A 1502 bp fragment partial ICS cDNA was amplified from pGEM-T vector through PCR using a set of primers. A binary vector, pGSA 1l31, was isolated from E. coli XLI Blue cells and was linearized using BamH 1 restriction endonuclease at the BamH 1 restriction site. Both cDNA and pGSA 1131 plasmid were subjected to end-blunting procedure. The partial ICS cDNA was ligated into pGSA 1131 binary vector through blunt end ligation to form a recombinant vector. Ligation mixture was transformed into competent E. coli XLI Blue cells via heat shock method. Single colonies were grown from a LB agar plate and five colonies were chosen and were heat disrupted to release their plasmid content. Colony PCR technique was performed using primers designed from conserved ICS domain. An amplified 800 bp fragment of the internal confirms the presence of the partial ICS cDNA in the colonies. Universiti Malaysia Sarawak, (UNIMAS) 2012 Final Year Project Report / IMRAD NonPeerReviewed text en http://ir.unimas.my/id/eprint/8125/1/Construction%20of%20A%20Plant%20Transformation%20Vector%20Containing%20A%20Partial%20Isochorismate%20Synthase%20cDNA%20%2824pgs%29.pdf text en http://ir.unimas.my/id/eprint/8125/8/Tai%20Ker%20Shing.pdf Tai, Ker Shing (2012) Construction of a plant transformation vector containing a partial isochorismate syhthase cDNA. [Final Year Project Report / IMRAD] (Unpublished) |
| spellingShingle | Q Science (General) Tai, Ker Shing Construction of a plant transformation vector containing a partial isochorismate syhthase cDNA |
| title | Construction of a plant transformation vector containing a partial isochorismate syhthase cDNA |
| title_full | Construction of a plant transformation vector containing a partial isochorismate syhthase cDNA |
| title_fullStr | Construction of a plant transformation vector containing a partial isochorismate syhthase cDNA |
| title_full_unstemmed | Construction of a plant transformation vector containing a partial isochorismate syhthase cDNA |
| title_short | Construction of a plant transformation vector containing a partial isochorismate syhthase cDNA |
| title_sort | construction of a plant transformation vector containing a partial isochorismate syhthase cdna |
| topic | Q Science (General) |
| url | http://ir.unimas.my/id/eprint/8125/ http://ir.unimas.my/id/eprint/8125/1/Construction%20of%20A%20Plant%20Transformation%20Vector%20Containing%20A%20Partial%20Isochorismate%20Synthase%20cDNA%20%2824pgs%29.pdf http://ir.unimas.my/id/eprint/8125/8/Tai%20Ker%20Shing.pdf |