Part:BBa_K564013

Upstream mutated chitoporin part fused with lacZ

ChiP encodes enterobacterial chitoporin required for uptake of chitin-derived oligosaccharides. This outer membrane porin is required for Salmonella and E.coli growth on chitooligosaccharides as a sole source of both carbon and nitrogen. It is only synthesized in the presence of these substrates. In nature it is regulated by sRNAs. In this biobrick we altered the ribosome binding site for ChiP regulator – ChiX (AAGAGG-->AGGAGA). It was done in order to show that the regulatory system depends on the specific base pairing between ChiP and its regulator. The promoter and reporter have not been altered compared to K564012.

This was verified by β-gal assays as shown in the table below. The measurements are in Miller Units

This system will not work in a strain deficient in hfq.

Strain	[http://ecoliwiki.net/colipedia/index.php/NM522 NM522]	[http://2011.igem.org/Team:DTU-Denmark/Project_testing_sRNA#Strain_construction IG302]
Genotype	chiX +	chiX -
PchiP(original):lacZ, BBa_K564012	467	4781
PchiP(mutated):lacZ, BBa_K564013	2353	2403

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
  COMPATIBLE WITH RFC[1000]