Part:BBa_K3122007

Outer membrane protein T (Vibrio cholerae) (TypeIIS adapted)

Vibrio cholerae’s ompT gene sequence was adapted to Golden Gate and MoClo assembly system, and expressed in this construct. The aim of it is to be the target for aptamers in the SELEX process, until developing a specific library against it. Unfortunately, we have not been able to clone this part for the moment. We expect future iGEM teams to resume our work from this point.

Biology

OmpT is described as a member of the family of outer membrane proteases called omptins (L. Vandeputte-Rutten, 2001). Moreover, other studies show a correlation between this protein and the regulation of cholera toxins, such as ToxR, and the presence of OmpT is directly connected to the sensitivity of the pathogen to host bile, hindering the colonization by the pathogen (E. Krukonis, 2003).

Characterization

Characterization of this part was meant to be conducted by building a transcriptional unit BBa_K3122013 and performing western blotting assays. Unfortunately, no results were obtained.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 227
  Illegal SpeI site found at 733
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 227
  Illegal NheI site found at 499
  Illegal SpeI site found at 733
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 227
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 227
  Illegal SpeI site found at 733
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 227
  Illegal SpeI site found at 733
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

L. Vandeputte-Rutten, R. Kramer, J. Kroon, N. Dekker, M. Egmond and P. Gros, "Crystal structure of the outer membrane protease OmpT from Escherichia coli suggests a novel catalytic site", The EMBO Journal, vol. 20, no. 18, pp. 5033-5039, 2001. Available: 10.1093/emboj/20.18.5033 [Accessed 19 October 2019].

E. Krukonis and V. DiRita, "From motility to virulence: sensing and responding to environmental signals in Vibrio cholerae", Current Opinion in Microbiology, vol. 6, no. 2, pp. 186-190, 2003. Available: 10.1016/s1369-5274(03)00032-8 [Accessed 19 October 2019].