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Determination of the Termination Efficiency of the Transcription Terminator Using Different Fluorescent Profiles in Green Fluorescent Protein Mutants

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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 Tl 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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Authors and Affiliations

  1. Biochemical Systems Laboratory, The Institute of Physical and Chemical Research (RIKEN), Saitama, 351-0198, Japan

    Takahiko Nojima, Angela C. Lin, Teruo Fujii & Isao Endo

  2. Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Fukuoka, 819-0395, Japan

    Takahiko Nojima

  3. Institute of Industrial Science, The University of Tokyo, Tokyo, 153-8505, Japan

    Teruo Fujii

  4. Saitama Industrial Technology Center, Saitama, 333-0844, Japan

    Isao Endo

Authors
  1. Takahiko Nojima
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  2. Angela C. Lin
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  3. Teruo Fujii
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  4. Isao Endo
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Correspondence to Takahiko Nojima.

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Nojima, T., Lin, A.C., Fujii, T. et al. Determination of the Termination Efficiency of the Transcription Terminator Using Different Fluorescent Profiles in Green Fluorescent Protein Mutants. ANAL. SCI. 21, 1479–1481 (2005). https://doi.org/10.2116/analsci.21.1479

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  • DOI: https://doi.org/10.2116/analsci.21.1479

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