Engineering a Yellow Thermostable Fluorescent Protein by Rational Design
Matthew R Anderson1, Caitlin M Padgett1, Cammi J Dargatz1
1Department of Chemistry and Biochemistry, Missouri State University, 901 South National Ave, Springfield, Missouri65897, United States.
Abstract:
Thermal green protein (TGP) is an extremely stable, highly soluble synthetic green fluorescent protein. The quantum yield of TGP is lower than the closest related natural fluorescent protein, monomeric Azami-Green. We improved the thermal recovery of TGP through the introduction of a chromophore mutation, Q66E. Furthermore, we developed a yellow thermal protein (YTP) via mutation of histidine 193 to tyrosine. Incorporation of Q66E into YTP (YTP-E) improved chemostability and pH stability. Both YTP and YTP-E have superior thermostability compared to TGP or TGP-E. These proteins offer a new option for green or yellow fluorescence under harsh chemical or thermal conditions.
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