Analytical Sciences
Online ISSN : 1348-2246
Print ISSN : 0910-6340
Original Papers
Determination of the Termination Efficiency of the Transcription Terminator Using Different Fluorescent Profiles in Green Fluorescent Protein Mutants
Takahiko NOJIMAAngela C. LINTeruo FUJIIIsao ENDO
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JOURNALS FREE ACCESS

2005 Volume 21 Issue 12 Pages 1479-1481

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Abstract

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.

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© 2005 by The Japan Society for Analytical Chemistry
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