High-throughput screening of expressed dna libraries in...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C07H 21/02 (2006.01) C12N 1/14 (2006.01) C12N 15/00 (2006.01) C12N 15/10 (2006.01)

Patent

CA 2376552

The invention provides a method for the expression of exogenous DNA libraries in filamentous fungi. The fungi are capable of processing intron-containing eukaryotic genes, and also can carry out post-translational processing steps such as glycosylation and protein folding. The invention provides for the use of fungi with altered morphology, which permits high-throughput screening and directed molecular evolution of expressed proteins. The same transformed fungi may be used (as shown in figure 1) to produce larger quantities of protein for isolation, characterization, and application testing, and may be suitable for commercial production of the protein as well.

L'invention concerne une méthode destinée à l'expression de bibliothèques d'ADN exogène dans des champignons filamenteux. Ces champignons sont capables de traiter des gènes eucaryotes contenant un intron et permettent également d'effectuer des opérations de traitement post-traductionnel, telles qu'une glycosylation et un repliement de protéine. L'invention concerne en outre l'utilisation de champignons à morphologie modifiée permettant la réalisation d'un criblage à débit élevé et d'une évolution moléculaire dirigée de protéines exprimées. Ces champignons transformés peuvent être utilisés (voir figure 1) pour produire des quantités plus importantes d'une protéine en vue d'une isolation, d'une caractérisation et d'un essai d'application, et peuvent être destinés à la production commerciale de cette protéine.

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