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Title: Circular permutant GFP insertion folding reporters

Patent ·
OSTI ID:1019100

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.

Research Organization:
Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
Sponsoring Organization:
USDOE
DOE Contract Number:
FG02-98ER62647
Assignee:
Los Alamos National Security, LLC (Los Alamos, NM)
Patent Number(s):
7,960,144
Application Number:
US Patent Application 12/215,053
OSTI ID:
1019100
Country of Publication:
United States
Language:
English

References (2)

Testing the role of chain connectivity on the stability and structure of dihydrofolate reductase from E. coli : Fragment complementation and circular permutation reveal stable, alternatively folded forms journal January 2001
Rapid protein-folding assay using green fluorescent protein journal July 1999

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