Circular permutant GFP insertion folding reporters

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

2011-06-14

2011-06-14

Monshipouri, Maryam (Department: 1656)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

Reexamination Certificate

active

07960144

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:
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Iwakura et al., 2000; Nat. Struct. Biol. 7(7): 580.
Smith and Matthews, 2001, Protein Sci. 10(1): 116.
Arai et al., 2003, J. Mol. Biol. 328(1): 273.

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