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The bifunctional dihydrofolate reductase thymidylate synthase of Tetrahymena thermophila provides a tool for molecular and biotechnology applications
BACKGROUND: Dihydrofolate reductase (DHFR) and thymidylate synthase (TS) are crucial enzymes in DNA synthesis. In alveolata both enzymes are expressed as one bifunctional enzyme. RESULTS: Loss of this essential enzyme activities after successful allelic assortment of knock out alleles yields an auxo...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2006
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1435751/ https://www.ncbi.nlm.nih.gov/pubmed/16549005 http://dx.doi.org/10.1186/1472-6750-6-21 |
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author | Herrmann, Lutz Bockau, Ulrike Tiedtke, Arno Hartmann, Marcus WW Weide, Thomas |
author_facet | Herrmann, Lutz Bockau, Ulrike Tiedtke, Arno Hartmann, Marcus WW Weide, Thomas |
author_sort | Herrmann, Lutz |
collection | PubMed |
description | BACKGROUND: Dihydrofolate reductase (DHFR) and thymidylate synthase (TS) are crucial enzymes in DNA synthesis. In alveolata both enzymes are expressed as one bifunctional enzyme. RESULTS: Loss of this essential enzyme activities after successful allelic assortment of knock out alleles yields an auxotrophic marker in ciliates. Here the cloning, characterisation and functional analysis of Tetrahymena thermophila's DHFR-TS is presented. A first aspect of the presented work relates to destruction of DHFR-TS enzyme function in an alveolate thereby causing an auxotrophy for thymidine. A second aspect is to knock in an expression cassette encoding for a foreign gene with subsequent expression of the target protein. CONCLUSION: This system avoids the use of antibiotics or other drugs and therefore is of high interest for biotechnological applications. |
format | Text |
id | pubmed-1435751 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2006 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-14357512006-04-13 The bifunctional dihydrofolate reductase thymidylate synthase of Tetrahymena thermophila provides a tool for molecular and biotechnology applications Herrmann, Lutz Bockau, Ulrike Tiedtke, Arno Hartmann, Marcus WW Weide, Thomas BMC Biotechnol Research Article BACKGROUND: Dihydrofolate reductase (DHFR) and thymidylate synthase (TS) are crucial enzymes in DNA synthesis. In alveolata both enzymes are expressed as one bifunctional enzyme. RESULTS: Loss of this essential enzyme activities after successful allelic assortment of knock out alleles yields an auxotrophic marker in ciliates. Here the cloning, characterisation and functional analysis of Tetrahymena thermophila's DHFR-TS is presented. A first aspect of the presented work relates to destruction of DHFR-TS enzyme function in an alveolate thereby causing an auxotrophy for thymidine. A second aspect is to knock in an expression cassette encoding for a foreign gene with subsequent expression of the target protein. CONCLUSION: This system avoids the use of antibiotics or other drugs and therefore is of high interest for biotechnological applications. BioMed Central 2006-03-20 /pmc/articles/PMC1435751/ /pubmed/16549005 http://dx.doi.org/10.1186/1472-6750-6-21 Text en Copyright © 2006 Herrmann et al; licensee BioMed Central Ltd. |
spellingShingle | Research Article Herrmann, Lutz Bockau, Ulrike Tiedtke, Arno Hartmann, Marcus WW Weide, Thomas The bifunctional dihydrofolate reductase thymidylate synthase of Tetrahymena thermophila provides a tool for molecular and biotechnology applications |
title | The bifunctional dihydrofolate reductase thymidylate synthase of Tetrahymena thermophila provides a tool for molecular and biotechnology applications |
title_full | The bifunctional dihydrofolate reductase thymidylate synthase of Tetrahymena thermophila provides a tool for molecular and biotechnology applications |
title_fullStr | The bifunctional dihydrofolate reductase thymidylate synthase of Tetrahymena thermophila provides a tool for molecular and biotechnology applications |
title_full_unstemmed | The bifunctional dihydrofolate reductase thymidylate synthase of Tetrahymena thermophila provides a tool for molecular and biotechnology applications |
title_short | The bifunctional dihydrofolate reductase thymidylate synthase of Tetrahymena thermophila provides a tool for molecular and biotechnology applications |
title_sort | bifunctional dihydrofolate reductase thymidylate synthase of tetrahymena thermophila provides a tool for molecular and biotechnology applications |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1435751/ https://www.ncbi.nlm.nih.gov/pubmed/16549005 http://dx.doi.org/10.1186/1472-6750-6-21 |
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