GENETICALLY EXPANDED CELL FREE PROTEIN SYNTHESIS S

专利名称：GENETICALLY EXPANDED CELL FREE

PROTEIN SYNTHESIS SYSTEMS, METHODSAND KITS

发明人：ALFONTA, Lital,CHEMLA, Yonatan,OZER,

Eden

申请号：EP15745249.1申请日：20150617公开号：EP3310924A1公开日：20180425

摘要：This invention relates to methods of producing a rare amino acid- or non-natural amino acid-containing protein in a cell free protein synthesis system and kits foruse in and for accomplishing same. Specifically, the methods comprise the steps ofexpressing at least one orthogonal suppressor tRNA (o-tRNA)/aminoacyl-tRNAsynthetase (aaRS) pair or derivatives thereof specific for incorporation of a rare aminoacid- or non-natural amino acid in an E. coli organism; preparing a lysate of said E. coliorganism expressing said orthogonal suppressor tRNA (o-tRNA)/aminoacyl-tRNAsynthetase (aaRS) pair; and contacting said lysate with a template DNA containing amutant gene in which at least one amino acid codon at a given site of the protein-encoding gene has been mutated into an amber or ochre mutation and further providinga cognate rare amino acid or non-natural amino acid and other factors necessary forprotein synthesis; wherein protein synthesis occurs following said contact to produce aprotein containing said at least one rare amino acid or said non-natural amino acid. Kitsfor use are described, as well.

申请人：B.G. Negev Technologies and Applications Ltd., at Ben-Gurion University

地址：P.O.BOx 653 841005 Beer Sheva IL

国籍：IL

代理机构：Zvesper, Thomas

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