Part:BBa_K4242004

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

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. 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 yedQ gene increased the intracellular c-di-GMP concentration and resulted in the enhanced secretion of EPS. We design the expression vector “pYDT-Ptac-yedQ” in E.coli BL21 and Acidithiobacillus ferrooxidans to characterize its function.

Fig 1. The gene circuit of expression vector “pYDT-Ptac-yedQ”

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

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

By overexpressing the yedQ gene, we found that A. ferrooxidans and E. coli BL21 formed thicker biofilms. yedQ also showed the highest concentration of c-di-GMP in E. coli 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.