luciferase gene activity

Makino Hideshi h_makino at kpu.ac.jp
Sun Nov 19 01:30:56 EST 1995


In article <DHz40I.4HH at hkuxb.hku.hk>, tsang chiu suet margaret <csmtsang at hkusua.hku.hk> writes:
> 
> HI Netters,
> we are trying to clone a promoter fragment into promega'a 
> pGL3 luciferase reporter. Is anyone know whether the 
> luciferase will still be active if a fusion enzyme is 
> formed? ie, If there is appro. 20 amino acid fuse with the
> lucifase in  the 5' end? 

I also fused ap. 20 amino acids to 5' of Firefly lucifesase(FFluc.) as a signal peptide.
Then, fused FFluc. reduced enzymatic acivity to ap. 1/10 times less than unfused.
In this case, detection with Western blot is more better, I think.

FFluc. is very sensitive, but sometimes its activity is lost easy.

mkn
---------------------------------
MAKINO Hideshi
Dept. Biochem. Kyoto Pref. Univ.
Shimogamo, Kyoto JAPAN
h_makino at kpu.ac.jp
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