Reporter

Part:BBa_K567008

Designed by: Yunfeng Ruan and Chunying Li   Group: iGEM11_SJTU-BioX-Shanghai   (2011-09-29)
Revision as of 10:42, 5 October 2011 by Cathy zks (Talk | contribs)

PT7-Luc-2AGG

This biobrick is constructed by putting modified enzyme luciferase under promoter T7 and controlled by lac operator. 2 AGG codons and 2 GCG codons are inserted after the ATG start codon of wild type luciferase (BBa_I712019). Modified luciferase keeps the activity of converting luciferin into oxyluciferin, during which bioluminescence will emit. This part is one of the reporter genes to testify the influence of different number of rare codons in regulating protein biosynthesis. This part is used as a measurement to testify the function of LacI -Ptrc-tRNA(Arg) (BBa_K567001) or sulA promoter-tRNA(Arg) (BBa_K567002). Cell is cultured in 50ug/ml kanamycin 10ug/ml tetracycline LB liquid medium. When the OD600 of the culture reaches 0.3 IPTG is added to make the final concentration 0.5nM to induce the synthesis of tRNA. Ultrasonication is used to release the luciferase from the cell. Sonics ON 3 seconds, OFF 3 seconds, total ultrasonication time 3minutes. Amount of bioluminescence produced can be detected using luminometer.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 48
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 48
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 48
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 911


[edit]
Categories
//cds/enzyme
Parameters
chassisER2566
functionIPTG induced luciferase expression
ligandsIsopropyl β-D-1-Thiogalactopyranoside (IPTG)