Difference between revisions of "Part:BBa K4614201"

Latest revision as of 15:20, 12 October 2023

The coding sequence that can be expressed in E. coli and induce filamentation.

SulA is an endogenous cell division inhibitor present in Escherichia coli^[1]. It is transcriptionally regulated by LexA and its expression is rapidly induced upon DNA damage, effectively halting cytoplasmic division. The inhibition of cell division by SulA occurs through direct interaction with the cell division protein FtsZ. FtsZ is a homologue of tubulin that forms a Z-ring at the middle of the cell, where other division proteins associate to facilitate cytoplasmic division. The disruption of FtsZ ring formation by SulA leads to the formation of elongated filaments, allowing the cells to replicate DNA but unable to undergo division^[2]. In projects, SulA was efficiently induced under the T7 promoter to induce the filamentous phenotype of Escherichia coli.

Reference

[1]Schoemaker, J M et al. “Regulation of cell division in Escherichia coli: SOS induction and cellular location of the sulA protein, a key to lon-associated filamentation and death.” Journal of bacteriology vol. 158,2 (1984): 551-61. doi:10.1128/jb.158.2.551-561.1984

[2]Mukherjee, A et al. “Inhibition of FtsZ polymerization by SulA, an inhibitor of septation in Escherichia coli.” Proceedings of the National Academy of Sciences of the United States of America vol. 95,6 (1998): 2885-90. doi:10.1073/pnas.95.6.2885

CAU_China 2023 used this part in their filamentation module. We inserted it under the T7 promoter of the pET32 vector and transferred the recombinant plasmid into E. coli (BL21) for induction of SulA production and filamentation at specific time points using IPTG.

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
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 376

Usage and Biology

CAU_China 2023 used this part in their filamentation module. We inserted it under the T7 promoter of the pET32 vector and transferred the recombinant plasmid into E. coli (BL21) for induction of SulA production and filamentation at specific time points using IPTG.

Figure A represents the control group, while Figure B shows the results of the SulA-induced filamentation experiment. IPTG was added to a final concentration of 20 mg/ml and chassis had been induced for over 10 hours.

The length of most hyphae exceeds 20 microns, demonstrating the filamentation effect of SulA.

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4614201 short</partinfo>
-SulA is an endogenous cell division inhibitor present in Escherichia coli[1]. It is transcriptionally regulated by LexA and its expression is rapidly induced upon DNA damage, effectively halting cytoplasmic division. The inhibition of cell division by SulA occurs through direct interaction with the cell division protein FtsZ. FtsZ is a homologue of tubulin that forms a Z-ring at the middle of the cell, where other division proteins associate to facilitate cytoplasmic division. The disruption of FtsZ ring formation by SulA leads to the formation of elongated filaments, allowing the cells to replicate DNA but unable to undergo division[2]. In projects, SulA was efficiently induced under the T7 promoter to induce the filamentous phenotype of Escherichia coli.
+SulA is an endogenous cell division inhibitor present in <i>Escherichia coli</i><sup>[1]</sup>. It is transcriptionally regulated by LexA and its expression is rapidly induced upon DNA damage, effectively halting cytoplasmic division. The inhibition of cell division by SulA occurs through direct interaction with the cell division protein FtsZ. FtsZ is a homologue of tubulin that forms a Z-ring at the middle of the cell, where other division proteins associate to facilitate cytoplasmic division. The disruption of FtsZ ring formation by SulA leads to the formation of elongated filaments, allowing the cells to replicate DNA but unable to undergo division<sup>[2]</sup>. In projects, SulA was efficiently induced under the T7 promoter to induce the filamentous phenotype of <i>Escherichia coli</i>.
+===Reference===
 [1]Schoemaker, J M et al. “Regulation of cell division in Escherichia coli: SOS induction and cellular location of the sulA protein, a key to lon-associated filamentation and death.” Journal of bacteriology vol. 158,2 (1984): 551-61. doi:10.1128/jb.158.2.551-561.1984
@@ Line 9: / Line 11: @@
 [2]Mukherjee, A et al. “Inhibition of FtsZ polymerization by SulA, an inhibitor of septation in Escherichia coli.” Proceedings of the National Academy of Sciences of the United States of America vol. 95,6 (1998): 2885-90. doi:10.1073/pnas.95.6.2885
-CAU_China 2023 used this part in their filamentation module. We inserted it under the T7 promoter of the pET32 vector and transferred the recombinant plasmid into E. coli (BL21) for induction of SulA production and filamentation at specific time points using IPTG.
+CAU_China 2023 used this part in their filamentation module. We inserted it under the T7 promoter of the pET32 vector and transferred the recombinant plasmid into <i>E. coli </i>(BL21) for induction of SulA production and filamentation at specific time points using IPTG.
 <!-- -->
 ===Sequence and Features===
@@ Line 20: / Line 22: @@
-CAU_China 2023 used this part in their filamentation module. We inserted it under the T7 promoter of the pET32 vector and transferred the recombinant plasmid into E. coli (BL21) for induction of SulA production and filamentation at specific time points using IPTG.
+CAU_China 2023 used this part in their filamentation module. We inserted it under the T7 promoter of the pET32 vector and transferred the recombinant plasmid into <i>E. coli</i> (BL21) for induction of SulA production and filamentation at specific time points using IPTG.
 https://static.igem.wiki/teams/4614/wiki/wiki-sh/wiki-safety/sh-sula.png