An activity-independent selection system of thermostable protein variants.
We describe an activity-independent method for the selection of thermostable mutants of any protein. It is based on a fusion construct comprising the protein of interest and a thermostable antibiotic resistance reporter, in such a way that thermostable mutants provide increased resistance in a thermophile. We isolated thermostable mutants of three human interferons and of two enzymes to demonstrate the applicability of the system.
Chautard
Hélène
H
Biométhodes SA, Genavenir 8, 5 rue Henri Desbruères, 91030 Evry, France.
Blas-Galindo
Emilio
E
Menguy
Thierry
T
Grand'Moursel
Laure
L
Cava
Felipe
F
Berenguer
José
J
Delcourt
Marc
M
D019943:Amino Acid Substitution / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics; D005560:Formate Dehydrogenases / Q000737:chemistry / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D016898:Interferon-alpha / Q000737:chemistry / Q000235:genetics; D016899:Interferon-beta / Q000737:chemistry / Q000235:genetics; D007371:Interferon-gamma / Q000737:chemistry / Q000235:genetics; D007612:Kanamycin / Q000494:pharmacology; D008049:Lipase / Q000737:chemistry / Q000235:genetics; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016963:Thermus thermophilus / Q000187:drug effects / Q000235:genetics / Q000254:growth & development
D016898:Interferon-alpha; D010446:Peptide Fragments; D011506:Proteins; D011994:Recombinant Proteins; D007612:Kanamycin; D016899:Interferon-beta; D007371:Interferon-gamma; D005560:Formate Dehydrogenases; D009713:Nucleotidyltransferases; C024251:kanamycin nucleotidyltransferase; D008049:Lipase