Part:BBa_K3376007

ldhp-AQP-Tr-tpxp-GFP-Tr/pSB1C3

Aquaporin is a transmembrane protein with the ability to facilitate bidirectional diffusion of water and H2O2. The DNA sequence was optimized for S. mutans UA159 and synthesized by Integrated DNA Technologies (IDT), Inc. In addition to Streptococcus cristatus, the gene function works well in E. coli as demonstrated in our work and consistent with the result in a paper by Huichun Tong, et al (J. Biol. Chem. (2019) 294(12) 4583–4595).

H2O2 Transporter

The activity of H2O2-regulated promoter of thiol peroxidase (tpxp) was enhanced in response to H2O2 only in the presence of the aquaporin protein, indicating AQP facilitates H2O2 permeation.

Regulation

The tpxp-GFP-Tr/pSB1C3 [BBa_K3376002] and ldhp-AQP-Tr-tpxp-GFP-Tr/pSB1C3 [BBa_K3376007] were transformed into E. coli Nissle strain. Because of the limited H2O2 diffusion across the cell membrane, the tpxp activity was not changed in the presence of H2O2. In aid of expression of aquaporin (AQP) to facilitate H2O2 permeation, the tpxp-driven GFP levels were enhanced by the increasing amounts of H2O2 in a dose-dependent manner.

Reference

Tong H, Wang X, Dong Y, Hu Q, Zhao Z, Zhu Y, Dong L, Bai F, Dong X. A Streptococcus aquaporin acts as peroxiporin for efflux of cellular hydrogen peroxide and alleviation of oxidative stress. J Biol Chem. 2019 Mar 22;294(12):4583-4595.

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 BsaI.rc site found at 1653