公开(公告)号：US20030049686A1

公开(公告)日：2003-03-13

申请号：US10112455

申请日：2002-03-29

Applicant: Korea Advanced Institute of Science and Technology

Inventor： Hak-Sung Kim ,  Geun-Joong Kim ,  Young-Hoon Cheon ,  Dong-Eun Lee

IPC: C12Q001/68 ,  G01N033/53 ,  C12P019/34 ,  C12P021/02 ,  C12N005/06 ,  C07K014/00

CPC classification number: C12N15/1058 ,  C07K14/43595 ,  C12N9/86 ,  C12N15/102

Abstract: The present invention relates to a method for manufacturing a mutant library of proteins with various sizes and sequences from a parental protein, microorganisms transformed with plasmids containing recombinant DNAs prepared by the insertion of a genomic DNA fragment into a defective template, a process for preparing proteins with different sizes and sequences from the parental protein which comprises the steps of culturing the transformed microorganisms and obtaining desired proteins from the culture, and proteins prepared by the said process. In accordance with the invention, a mutant library of proteins with various sizes and sequences can be manufactured from a parental protein in an efficient and simple manner, by constructing a library of microorganisms transformed with recombinant plasmids containing E. coli genomic DNA fragments inserted into defective genes and selecting clones expressing proteins with restored functions or modified characteristics.

Abstract translation: 本发明涉及一种用于制备来自亲本蛋白质的各种大小和序列的蛋白质突变文库的方法，用含有通过将基因组DNA片段插入缺陷模板制备的重组DNA的质粒转化的微生物，制备蛋白质的方法 其具有来自亲本蛋白的不同大小和序列，其包括培养转化的微生物并从培养物获得所需蛋白质的步骤，以及通过所述方法制备的蛋白质。 根据本发明，可以通过构建用包含插入有缺陷的大肠杆菌基因组DNA片段的重组质粒转化的微生物文库，以有效和简单的方式从亲本蛋白质制造出具有不同大小和序列的蛋白质的突变文库 基因并选择表达具有恢复功能或修饰特征的蛋白质的克隆。