Difference between revisions of "Part:BBa K4242004"

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<center><b>Fig. 1.</b> The quantification of intracellular c-di-GMP in C. testosteroni WDL7/pYedQ2, the wild-type strain and WDL7/YhjH. nd, not determined (a). Crystal violet staining of WDL7/pYedQ2, WDL7 and WDL7/YhjH strains for quantifying relative biofilm biomass (b). * indicates significantly difference when compared with the WDL7 wild type strain, * p < 0.05, *** p < 0.001[1].</center>
 
<center><b>Fig. 1.</b> The quantification of intracellular c-di-GMP in C. testosteroni WDL7/pYedQ2, the wild-type strain and WDL7/YhjH. nd, not determined (a). Crystal violet staining of WDL7/pYedQ2, WDL7 and WDL7/YhjH strains for quantifying relative biofilm biomass (b). * indicates significantly difference when compared with the WDL7 wild type strain, * p < 0.05, *** p < 0.001[1].</center>
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= CUG-China 2024--Contribution =
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YedQ is a diguanylate cyclase from <i>Escherichia coli.<i> It is registered in 2022 by CUG-China, which works with the phosphodiesterase (YhjH) modulated the in vivo c-di-GMP levels.
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The expression of the <i>yedQ<i> gene increased the intracellular c-di-GMP concentration and resulted in the enhanced secretion of EPS. We design the expression vector “pYDT-<i>Ptac<i>-<i>yedQ<i>” in <i>E.coli<i> BL21 and <i>Acidithiobacillus ferrooxidans<i> to characterize its function.
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<center>https://static.igem.wiki/teams/5323/parts/c1.png</center>
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<center>Fig 1. The gene circuit of expression vector “pYDT-<i>Ptac<i>-<i>yedQ<i>”
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We examined the role of <i>yedQ<i> by overexpressing <i>yedQ<i> and measuring biofilm formation in overexpressing engineered bacteria versus c-di-GMP content in bacteria.
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<center>https://static.igem.wiki/teams/5323/parts/c2.png</center>
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<center>Fig 2.(A) <i>Acidithiobacillus ferrooxidans<i> pore plate biofilm formation column.(B) <i>Escherichia coli<i>  pore plate biofilm formation column.(C) Schematic diagram of c-di-GMP content of engineered <i>E.coli<i> BL21 pYDT-0053, pYDT-1379, pYDT-1373, pYDT-yedQ. (D) Indicates the standard curve of c-di-GMP.
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By overexpressing the <i>yedQ<i> gene, we found that <i>A. ferrooxidans<i> and <i>E. coli BL21<i> formed thicker biofilms. <i>yedQ<i> also showed the highest concentration of c-di-GMP in <i>E. coli<i> BL21 strains, suggesting that high levels of c-di-GMP can regulate the formation of thicker biofilms.
  
 
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Revision as of 08:45, 1 October 2024


yedQ

This part sequence originated from Escherichia coli encodes YedQ protein. YedQ is diguanylate cyclase.

Usage and Biology

YedQ can work with the phosphodiesterase (YhjH) modulated the in vivo c-di-GMP levels. The expression of the yedQ gene increased the intracellular c-di-GMP concentration and resulted in the enhanced secretion of EPS[1]. So, The protein YedQ can promote bacterial self-aggregation and adhesion onto negatively charged surfaces. In E. coli, extracellular DNA (eDNA) was increased as expected for the deletions of yedQ. As a result of the significantly enhanced motility, but contrary to current models of decreased biofilm formation with decreased diguanylate cyclase activity, early biofilm formation increased dramatically for the deletions of yedQ[2].

The quantification of intracellular c-di-GMP

YedQ.png
Fig. 1. The quantification of intracellular c-di-GMP in C. testosteroni WDL7/pYedQ2, the wild-type strain and WDL7/YhjH. nd, not determined (a). Crystal violet staining of WDL7/pYedQ2, WDL7 and WDL7/YhjH strains for quantifying relative biofilm biomass (b). * indicates significantly difference when compared with the WDL7 wild type strain, * p < 0.05, *** p < 0.001[1].

CUG-China 2024--Contribution

YedQ is a diguanylate cyclase from Escherichia coli.<i> It is registered in 2022 by CUG-China, which works with the phosphodiesterase (YhjH) modulated the in vivo c-di-GMP levels. The expression of the <i>yedQ<i> gene increased the intracellular c-di-GMP concentration and resulted in the enhanced secretion of EPS. We design the expression vector “pYDT-<i>Ptac<i>-<i>yedQ<i>” in <i>E.coli<i> BL21 and <i>Acidithiobacillus ferrooxidans<i> to characterize its function.

c1.png
Fig 1. The gene circuit of expression vector “pYDT-<i>Ptac<i>-<i>yedQ<i>”

We examined the role of <i>yedQ<i> by overexpressing <i>yedQ<i> and measuring biofilm formation in overexpressing engineered bacteria versus c-di-GMP content in bacteria.

<center>c2.png
Fig 2.(A) <i>Acidithiobacillus ferrooxidans<i> pore plate biofilm formation column.(B) <i>Escherichia coli<i> pore plate biofilm formation column.(C) Schematic diagram of c-di-GMP content of engineered <i>E.coli<i> BL21 pYDT-0053, pYDT-1379, pYDT-1373, pYDT-yedQ. (D) Indicates the standard curve of c-di-GMP.

By overexpressing the <i>yedQ<i> gene, we found that <i>A. ferrooxidans<i> and <i>E. coli BL21<i> formed thicker biofilms. <i>yedQ<i> also showed the highest concentration of c-di-GMP in <i>E. coli<i> BL21 strains, suggesting that high levels of c-di-GMP can regulate the formation of thicker biofilms.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1591
  • 1000
    COMPATIBLE WITH RFC[1000]

References

[1] Yang S, Wu Y, Qu C, et al. Quantitative analysis of the surficial and adhesion properties of the Gram-negative bacterial species Comamonas testosteroni modulated by c-di-GMP[J]. Colloids Surf B Biointerfaces, 2021,198:111497.

[2] Sanchez-Torres V, Hu H, Wood T K. GGDEF proteins YeaI, YedQ, and YfiN reduce early biofilm formation and swimming motility in Escherichia coli[J]. Appl Microbiol Biotechnol, 2011,90(2):651-658.