公开(公告)号：US06562622B1

公开(公告)日：2003-05-13

申请号：US09674677

申请日：2000-12-11

申请人： Gregory Coia ,  Peter John Hudson ,  Peter Iliades ,  Robert Alexander Irving

发明人： Gregory Coia ,  Peter John Hudson ,  Peter Iliades ,  Robert Alexander Irving

IPC分类号： C12N1500

CPC分类号： C12N9/127 ,  C07K16/082 ,  C07K16/18 ,  C07K2317/622 ,  C12N15/102 ,  C12N15/1041

摘要： The present invention provides a method for the mutation, synthesis and selection of a protein which binds to a target molecule, the method comprising: (a) incubating a replicable mRNA molecule encoding the protein with ribonucleoside triphosphate precursors of RNA and an RNA-directed RNA polymerase, wherein the RNA-directed RNA polymerase replicates the mRNA molecule but introduces mutations thereby generating a population of mutant mRNA molecules; (b) incubating the mutant mRNA molecules from step (a) with a translation system under conditions which results in the synthesis of population of mutant proteins such that after translation, mutant proteins are linked to their encoding mRNA molecules thereby forming a population of mutant proteins such that after translation, mutant proteins are linked to their encoding mRNA molecules thereby forming a population of mutant/mRNA complexes; (c) selecting one or more mutant protein/mRNA complex(es) by exposing the population of mutant protein/mRNA complexes from step (b) to the target molecule and recovering the mutant protein/mRNA complex(es) bound thereto.

摘要翻译： 本发明提供了与靶分子结合的蛋白质的突变，合成和选择的方法，所述方法包括：（a）将编码该蛋白的可复制mRNA分子与RNA的核糖核苷三磷酸前体和RNA指导的RNA 聚合酶，其中RNA指导的RNA聚合酶复制mRNA分子，但引入突变，从而产生突变体mRNA分子群; （b）在导致突变蛋白质群体合成的条件下将来自步骤（a）的突变体mRNA分子与翻译系统一起孵育，使得在翻译后，突变蛋白质与其编码mRNA分子连接，从而形成突变蛋白质群 使得翻译后，突变蛋白与其编码mRNA分子连接，从而形成突变体/ mRNA复合物群; （c）通过将来自步骤（b）的突变蛋白/ mRNA复合物的群体暴露于靶分子并回收与其结合的突变蛋白/ mRNA复合物来选择一种或多种突变蛋白/ mRNA复合物。