Part:BBa_K1036000

lux pL controlled luxR with lux pR controlled ndh (LVA-tag) coding for NADH dehydrogenase II

Description

This part is made up of a quorum sensing promoter and ndh gene with a LAV-tag. Under control of the quorum sensing promoter, ndh gene will express periodically as the promoter is periodically activated and repressed. Since the ndh gene is described detailed in Part: BBa_K1036001https://parts.igem.org/Part:BBa_K1036001, here is a detailed description of quorum sensing promoter. Firstly, we should be familiar with the quorum sensing mechanism, the luxI gene is at low expression level and produces LuxI protein that synthesizes a kind of acyl-homoserine lactone (AHL), which is a small molecule that can diffuse across the cell membrane and mediate intercellular coupling when it reaches the threshold as enough biomass accumulated. AHL will bind intracellular protein LuxR, which is also consecutively produced by luxR gene. The LuxR-AHL complex can activate the luxI promoter, and the positive feedback loop is built. However, since in this circuit the luxI gene behind the luxI promoter is changed into ndh gene (with LVA-tag), so it could only be activated by AHL produced by luxI gene on another plasmid, which contains Part: BBa_K1036003.

Protocol

Ndh with LVA tag is formed as a composite by the ligation of quorum sensing promoter luxR via digestion and ligation.

Experimental Data

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2544
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1101