Patent ID: 9637528
Date: 2017-05-02
CPC Classifications: C07K,C12N,G01N

Claim:
1. A method of generating a polynucleotide encoding an enhanced folding variant of a cyan fluorescent protein, comprising: (a) providing a polynucleotide encoding a fusion protein comprising a heterologous folding interference domain inserted into a permissive site of a cyan fluorescent protein, wherein (b) providing a library of test polynucleotides encoding mutants of (a), wherein the library of test polynucleotides is obtained from the polynucleotide of (a) mutagenized by error-prone PCR mutagenesis; (c) expressing the library of test polynucleotides to produce the mutants of the fusion protein; (d) assaying fluorescence of the mutants of the fusion protein; (e) selecting from the library a test polynucleotide encoding a mutant of the fusion protein with increased fluorescence compared to the fluorescence of the fusion protein; and (f) removing the polynucleotide encoding the heterologous folding interference domain from the selected test polynucleotide of step (e) to generate a polynucleotide encoding an enhanced folding variant of the cyan fluorescent protein.