Part:BBa_K389004

Luciferase from pGL4.10[luc2]

Structure of Photinus pyralis firefly luciferase. Image taken from: http://en.wikipedia.org/wiki/Luciferase.

A firefly (Photinus pyralis) luciferase from Promega's pGL4.10[luc2] vector. Luciferase is a sensitive reporter gene.

Usage and Biology

The activity of a luciferase is measured in a luminometer - so only use this BioBrick if you have one in your lab.

Firefly luciferase oxidizes its substrate, firefly luciferin, in presence of ATP and oxygen, emitting light ([http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VSR-4CJB7RR-7&_user=2459438&_coverDate=03%2F31%2F1996&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_acct=C000057302&_version=1&_urlVersion=0&_userid=2459438&md5=ac60aa60eaad17b2aa1c4b31f954d0a8&searchtype=a Conti et al., 1996]). Because the firefly luciferin does not pass the cell membrane of E. coli under "normal" cultivation conditions the cells have to be lysated before the luciferase activity measurement.

Important parameters

Experiment	Result
Behaviour during cultivation	production is growth dependent degradation in stationary growth phase
Kinetic of luciferin conversion	max. output between 20 - 40 s
Limit of detection (LOD)	162 RLU ~ 0.3 % of BBa_J23103 output
Limit of quantification (LOQ)	306 RLU ~ 0.7 % of BBa_J23103 output

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 NgoMIV site found at 64
  Illegal NgoMIV site found at 1408
  Illegal NgoMIV site found at 1429
  Illegal AgeI site found at 1132
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1314