Method for constructing oligodesoxyribonucleotide binomial...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/00 (2006.01) B01J 19/00 (2006.01) C07B 61/00 (2006.01) C07H 21/00 (2006.01)

Patent

CA 2391999

The invention relates to a mutagenesis method for constructing oligodesoxyribonucleotide binomial libraries with mutagenized codon involving the utilization of two sets of desoxynucleoside-phosphoramitides that are protected in the (5ï) hydroxyl by the orthogonal protection groups which are combined during oligodesoxyribonucleotide synthesis. Said method is a valuable tool for the study of the structure-function relation of proteins and engineering of the latter since few replacements of amino acids by proteins can be generated in a controlled and predictable manner, thereby making it possible to investigate the individual importance of each of the wild amino acids for the protein function while preventing their functional destruction with the purpose of improving innumerable amounts of proteins with commercial applications, for instance, the enzymes called subtilisins which are used in biodegradable detergents.

L'invention concerne un procédé de mutagenèse ayant pour objectif la construction de banques binomiales d'oligodésoxyribonucléotides mutagénisés au niveau du codon consistant à utiliser deux ensembles de désoxynucléoside-phosphoramidites protégés dans l'hydroxyle 5' à l'aide de groupes protecteurs orthogonaux, lesquels sont combinés lors de la synthèse des oligodésoxyribonucléotides. Ce procédé constitue un outil précieux pour l'étude du rapport entre la structure et la fonction des protéines et le génie protéique associé, étant donné que peu de substituts d'acides aminés par protéine sont générés d'une manière régulée et prévisible. Ainsi, on peut étudier l'importance individuelle de chaque acide aminé sylvestre en termes de fonction protéique tout en évitant leur destruction fonctionnelle dans le but d'améliorer une infinité de protéines à application commerciale, comme les enzymes appelées subtilisines, par exemple, que l'on trouve dans la composition des détergents biodégradables.

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