Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Yunfeng Ruan and Chunying Li
Date created: 2011-09-28 11:00:00
Date modified: 2015-05-08 01:12:43
PT7-Luc-4AGG
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_K567009_sequence (Version 1) |
Description
this biobrick is constructed by putting modified enzyme luciferase under promoter T7 and controlled by lac operator. 4 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 -Ptac-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.