Creation of circularly permutated yellow fluorescent proteins using fluorescence screening and a tandem fusion template

Biotechnology Letters
Jason Jui-Hsuan ChiangKevin Truong

Abstract

By experimenting with many different circularly permutated yellow fluorescent protein (cpYFP) variants as acceptors in fluorescence resonance energy transfer based biosensors, the optimal dynamic range can be discovered by sampling the possibilities of relative fluorophore orientations before and after bioactivity. Hence, to facilitate the sampling process, we introduced a new approach to construct a library of cpYFP variants using fluorescence screening and a tandem fusion template. This new approach is rapid because it does not require creating intermediate N- and C-terminal fragments and it allows quick screening for positive colonies by fluorescence. As a demonstration, eleven cpYFP variants were created and eight showed fluorescence. The emission and excitation spectra of these cpYFP variants showed strong similarity to YFP and therefore can be used in replacement.

References

Sep 29, 1999·Proceedings of the National Academy of Sciences of the United States of America·G S BairdR Y Tsien
Mar 15, 2001·Proceedings of the National Academy of Sciences of the United States of America·T NagaiA Miyawaki
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Jul 13, 2004·Proceedings of the National Academy of Sciences of the United States of America·Takeharu NagaiAtsushi Miyawaki

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Citations

Jun 8, 2011·PloS One·Bo ShuiMichael I Kotlikoff
Oct 15, 2010·Protein Science : a Publication of the Protein Society·Xi ChenKevin Truong
Aug 30, 2012·Angewandte Chemie·Jing WangChuan He
Aug 30, 2019·International Journal of Molecular Sciences·Alexander I KostyukDmitry S Bilan
Nov 17, 2010·Journal of the American Chemical Society·Boxuan Simen ZhaoPeng R Chen

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