Difference between revisions of "Part:BBa K3861003"

Revision as of 14:49, 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 (<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.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K3861003 short</partinfo>
+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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