Difference between revisions of "Part:BBa K3861003"
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The coding region of the Thymidylate synthase (ThyA, EC2.1.1.45). The enzyme catalyzes the reductive methylation of 2'-deoxyuridine-5'-monophosphate (dUMP) to 2'-deoxythymidine-5'-monophosphate (dTMP). <i>Salmonella</i> typhimurium <i>thyA</i> knock out mutants cannot synthesize DNA if no thymidine monophosphate (dTMP) is available to the cells (Neuhard and Kelln, 1996). Furthermore, <i>thyA</i> mutants were shown to not grow intercellular in macrophage-like and human epithelial cell lines <i>in vitro</i> (Kok et al., 2001). Thus, for dTMP auxotrophic bacterial strains can be created. We used this knowledge to our advantage and used <i>thyA</i> as selection marker. By deleting the chromosomal <i>thyA</i> gene and reintroducing it with one of our plasmids (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K3861005">pKF01</a>), we effectively bypassed the need for an antibiotic based selection marker system. | The coding region of the Thymidylate synthase (ThyA, EC2.1.1.45). The enzyme catalyzes the reductive methylation of 2'-deoxyuridine-5'-monophosphate (dUMP) to 2'-deoxythymidine-5'-monophosphate (dTMP). <i>Salmonella</i> typhimurium <i>thyA</i> knock out mutants cannot synthesize DNA if no thymidine monophosphate (dTMP) is available to the cells (Neuhard and Kelln, 1996). Furthermore, <i>thyA</i> mutants were shown to not grow intercellular in macrophage-like and human epithelial cell lines <i>in vitro</i> (Kok et al., 2001). Thus, for dTMP auxotrophic bacterial strains can be created. We used this knowledge to our advantage and used <i>thyA</i> as selection marker. By deleting the chromosomal <i>thyA</i> gene and reintroducing it with one of our plasmids (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K3861005">pKF01</a>), we effectively bypassed the need for an antibiotic based selection marker system. | ||
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Revision as of 14:50, 17 October 2021
ThyA CDS
The coding region of the Thymidylate synthase (ThyA, EC2.1.1.45). The enzyme catalyzes the reductive methylation of 2'-deoxyuridine-5'-monophosphate (dUMP) to 2'-deoxythymidine-5'-monophosphate (dTMP). Salmonella typhimurium thyA knock out mutants cannot synthesize DNA if no thymidine monophosphate (dTMP) is available to the cells (Neuhard and Kelln, 1996). Furthermore, thyA mutants were shown to not grow intercellular in macrophage-like and human epithelial cell lines in vitro (Kok et al., 2001). Thus, for dTMP auxotrophic bacterial strains can be created. We used this knowledge to our advantage and used thyA as selection marker. By deleting the chromosomal thyA gene and reintroducing it with one of our plasmids (pKF01), we effectively bypassed the need for an antibiotic based selection marker system.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]