首页> 外国专利> Novel method for identification of clones with a desired biological property, starting from an expression, gene bank (2001 / 06)

Novel method for identification of clones with a desired biological property, starting from an expression, gene bank (2001 / 06)

机译:从表达基因库开始鉴定具有所需生物学特性的克隆的新方法(2001/06)

摘要

The present invention relates to a novel method for the identification and/or characterization of clones conferring a desired biological property from an expression library. The method of the invention comprises the step of analyzing for the expression of at least one (poly)peptide, such as a tag expressed as a fusion protein, together with a recombinant insert of a clone of said expression library, wherein the clones of said expression library are arranged in arrayed form. Said (poly)peptide may be fused N-terminally or C-terminally to said insert. The method of the invention further comprises the steps of contacting a ligand specifically interacting with a (poly)peptide expressed by the insert of a clone conferring said desired biological property with a first replica of said library of clones in arrayed form and analyzing said library of clones for the occurrence of an interaction, and/or carrying out a hybridization or an oligonucleotide fingerprint with a nucleic acid probe specific for the insert of a clone conferring said desired biological property with a second replica of said library of clones arranged in arrayed form and analyzing said library of clones for the occurrence of a specific hybridization. Finally, the method of the invention requires the identification of clones wherein an expression of the at least one (poly)peptide in step (a) and/or an interaction in step (b) and/or a hybridization or an oligonucleotide fingerprint in step (c) can be detected. The present invention also relates to a kit useful for carrying out the method of the invention.
机译:本发明涉及从表达文库鉴定和/或鉴定赋予所需生物学特性的克隆的新方法。本发明的方法包括以下步骤:分析至少一种(多)肽,例如表达为融合蛋白的标签,以及所述表达文库的克隆的重组插入片段的表达,其中表达式库以阵列形式排列。所述(多肽)肽可以在N末端或C末端与所述插入物融合。本发明的方法进一步包括以下步骤:使与赋予所述所需生物学特性的克隆的插入物表达的(多)肽特异性相互作用的配体与所述克隆文库的第一复制品呈阵列形式接触,并分析所述cDNA文库。用于发生相互作用的克隆,和/或与特定于克隆插入物的核酸探针进行杂交或寡核苷酸指纹分析,所述核酸探针赋予所述期望的生物学特性,所述克隆的第二个副本以阵列形式排列,并分析所述克隆文库中是否发生特异性杂交。最后,本发明的方法需要鉴定其中在步骤(a)中表达至少一种(多)肽和/或在步骤(b)中相互作用和/或在步骤中杂交或寡核苷酸指纹的克隆。 (c)可以被检测到。本发明还涉及用于实施本发明方法的试剂盒。

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