Circular permutant GFP insertion folding reporters

Details Circular permutant GFP insertion folding reporters Circular permutant GFP insertion folding reporters

2008-06-24

2008-06-24

Monshipouri, Maryam (Department: 1656)

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Oxidoreductase

C435S252300, C435S320100, C435S006120, C536S023100

Reexamination Certificate

active

07390640

ABSTRACT:
Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited.

REFERENCES:
patent: 6448087 (2002-09-01), Waldo
patent: 7271241 (2007-09-01), Waldo
Waldo and Cabantous, U.S. Appl. No. 10/973,693, filed Oct. 24, 2004.
Smith and Matthews, 2001, Protein Sci. 10(1): 116.
Arai et al., 2003, J. Mol. Biol. 328(1):273.

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