Difference between revisions of "Part:BBa K1040001"

Revision as of 14:16, 9 October 2023 (view source) Ruosu (Talk | contribs) m ← Older edit		Revision as of 14:18, 9 October 2023 (view source) Ruosu (Talk | contribs) m Newer edit →
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	===Improvement===		===Improvement===
−	<p>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 E. coli (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		https://static.igem.wiki/teams/4614/wiki/wiki-sh/wiki-safety/sh-sula.png
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	===References of CAU_China===		===References of CAU_China===
−	<p>[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/p>	+	[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/p>
−	<p>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.</p>	+	[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.</p>
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Revision as of 14:18, 9 October 2023

E.coli(K12) sulA

During cell division, FtsZ moves to the division site, and is essential for recruiting other proteins that produce a new cell wall between the dividing cells in E. coli .The Z-ring forms from smaller subunits of FtsZ filaments. SulA sequesters FtsZ, preventing the formation of the polymerized Z Ring . The function of SulA is also linked to stressors like DNA damage. DNA damage induces SulA. SulA binds to self-recognizing FtsZ sitesprevents the polymerization and GTPase activity of FtsZ. The cell can link DNA damage to inhibiting cell division.

Contribution From CAU_China 2023

Group: CAU_China, 2023 https://2023.igem.org/Team:CAU_China

Author: Yao muchen

Summary:We added some experimental conditions and result images

Biology

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.

Improvement

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.

References of CAU_China

[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/p>

[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.

Sequence and Features