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1. 发明申请

WO2005121351A2 METHODS FOR TRANSFORMING AND EXPRESSION SCREENING OF FILAMENTOUS FUNGAL CELLS WITH A DNA LIBRARY 审中-公开
标题翻译：用DNA文库对真菌真菌进行转化和表达筛选的方法
公开(公告)号：WO2005121351A2
公开(公告)日：2005-12-22
申请号：PCT/US2005018871
申请日：2005-05-26
申请人： NOVOZYMES INC
发明人： TETER SARAH , LAMSA MICHAEL , CHEERY JOEL , WARD CONNIE
IPC分类号： C12N1/16 , C12N15/74 , C12P21/06
CPC分类号： C12N15/1037 , C12N15/1086 , C12N15/80 , C40B40/02
摘要： The present invention relates to methods for expression screening of filamentous fungal transformants, comprising: (a) isolating single colony transformants of a DNA library introduced into E. coli; (b) preparing DNA from each of the single colony E. coli transformants; (c) introducing a sample of each of the DNA preparations of step (b) into separate suspensions of protoplasts of a filamentous fungus to obtain transformants thereof, wherein each transformant contains one or more copies of an individual polynucleotide from the DNA library; (d) growing the individual filamentous fungal transformants of step (c) on selective growth medium, thereby permitting growth of the filamentous fungal transformants, while suppressing growth of untransformed filamentous fungi; and (e) measuring activity or a property of each polypeptide encoded by the individual polynucleotides.
摘要翻译：本发明涉及丝状真菌转化体的表达筛选方法，其包括：（a）分离导入大肠杆菌的DNA文库的单个菌落转化体; （b）从每个单个菌落的大肠杆菌转化体制备DNA; （c）将步骤（b）的每种DNA制剂的样品引入到丝状真菌的原生质体的分离悬浮液中以获得其转化体，其中每个转化体含有来自DNA文库的单个多核苷酸的一个或多个拷贝; （d）在选择性生长培养基上生长步骤（c）的单个丝状真菌转化体，从而允许丝状真菌转化体生长，同时抑制未转化的丝状真菌的生长; 和（e）测量由各个多核苷酸编码的每种多肽的活性或性质。

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