Determination of the termination efficiency of the transcription terminator using different fluorescent profiles in green fluorescent protein mutants.
An approach in determining the intrinsic termination efficiency (%T) of transcription termination using green fluorescent protein (GFP) mutants was developed. This approach utilizes a cassette vector in which the tested terminator is introduced between two GFP mutant genes: an ultraviolet-optimized mutant (GFPuv: F99S, M153T, V163A) and a blue-shifted mutant (BFP: F64L, S65T, T145F). The ratio of the fluorescence intensity of BFP to GFPuv after transcription and translation represents the termination efficiency of the terminator. E. coli ribosomal RNA operon T1 terminator, phage lambda terminator site R2, E. coli tryptophane attenuater were introduced into the vector, and their transcriptional efficiencies were estimated as 89, 79, and 24%, respectively, showing good agreement with published data.
Nojima
Takahiko
T
Biochemical Systems Laboratory, The Institute of Physical and Chemical Research (RIKEN), Saitama 351-0198, Japan. nojimtcm@mbox.nc.kyushu-u.ac.jp
Lin
Angela C
AC
Fujii
Teruo
T
Endo
Isao
I
D001483:Base Sequence; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics*; D009154:Mutation; D009690:Nucleic Acid Conformation; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
D017931:DNA Primers; D049452:Green Fluorescent Proteins