A h c t New and highly effective strategies for directed enzyme evolution in vitro have been developed in the protein engineering field. They allow engineering all kinds of enzymes in vitro so that new ones with novel functions and features can be obtained by the methods of enorprone PCR , DNA shuffling ( exon shuffling) , hybrid enzyme, random-priming in vfiro recombination ( RPR) , stagger extension process ( StEP ) , random-directed mutagenesis in vitro, etc. , even with little knowlodge of spatial structure and catalytic mechanism of enzymes in advance. The process that would take millions of years in nature can in principle be accomplished in the test within several years.IT has been proved now not only that scientists can investigate structure-function of enzyme in unprecedented depth and range, but also that study and applied ranges of protein engineering have been greatly broadened since the techniques of directed evolution of enzyme in vitro were found recently. It is unnecessary for us to understand the spatial structure and catalytic mechanism in advance to engineer proteins or
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