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2. 发明专利

DE69620766T2 未知
公开(公告)号：DE69620766T2
公开(公告)日：2004-11-18
申请号：DE69620766
申请日：1996-08-12
申请人： NOVOZYMES AS
发明人： OKKELS SIGURD
IPC分类号： C12N15/09 , C12N1/19 , C12N9/20 , C12N15/10 , C12P21/02 , C12R1/865 , C12N15/62 , C12N15/63
摘要： The present invention relates to a method for preparing positive polypeptide variants by shuffling different nucleotide sequences of homologous DNA sequences by in vivo recombination comprising the steps of a) forming at least one circular plasmid comprising a DNA sequence encoding a polypeptide, b) opening said circular plasmid(s) within the DNA sequence(s) encoding the polypeptide(s), c) preparing at least one DNA fragment comprising a DNA sequence homologous to at least a part of the polypeptide coding region on at least one of the circular plasmid(s), d) introducing at least one of said opened plasmid(s), together with at least one of said homologous DNA fragment(s) covering full-length DNA sequences encoding said polypeptide(s) or parts thereof, into a recombination host cell, e) cultivating said recombination host cell, and f) screening for positive polypeptide variants, wherein more than one cycle of step a) to f) are performed and/or wherein the shuffling is carried out with 2 or more DNA fragments with partially overlapping regions.

3. 发明专利

DE69620766D1 未知
公开(公告)号：DE69620766D1
公开(公告)日：2002-05-23
申请号：DE69620766
申请日：1996-08-12
申请人： NOVOZYMES AS
发明人： OKKELS SIGURD
IPC分类号： C12N15/09 , C12N1/19 , C12N9/20 , C12N15/10 , C12P21/02 , C12R1/865 , C12N15/62 , C12N15/63
摘要： The present invention relates to a method for preparing positive polypeptide variants by shuffling different nucleotide sequences of homologous DNA sequences by in vivo recombination comprising the steps of a) forming at least one circular plasmid comprising a DNA sequence encoding a polypeptide, b) opening said circular plasmid(s) within the DNA sequence(s) encoding the polypeptide(s), c) preparing at least one DNA fragment comprising a DNA sequence homologous to at least a part of the polypeptide coding region on at least one of the circular plasmid(s), d) introducing at least one of said opened plasmid(s), together with at least one of said homologous DNA fragment(s) covering full-length DNA sequences encoding said polypeptide(s) or parts thereof, into a recombination host cell, e) cultivating said recombination host cell, and f) screening for positive polypeptide variants, wherein more than one cycle of step a) to f) are performed and/or wherein the shuffling is carried out with 2 or more DNA fragments with partially overlapping regions.

5. 发明专利

DE69632538D1 未知
公开(公告)号：DE69632538D1
公开(公告)日：2004-06-24
申请号：DE69632538
申请日：1996-08-12
申请人： NOVOZYMES AS
发明人： OKKELS SIGURD , SVENDSEN ALLAN , BORCH KIM , THELLERSEN MARIANNE , PATKAR ANANT , PETERSEN AABY , ROYER C , KRETZSCHMAR TITUS
IPC分类号： C12N15/09 , C11D3/386 , C12N9/20

6. 发明专利

DE69632538T2 未知
公开(公告)号：DE69632538T2
公开(公告)日：2005-05-19
申请号：DE69632538
申请日：1996-08-12
申请人： NOVOZYMES AS
发明人： OKKELS SIGURD , SVENDSEN ALLAN , BORCH KIM , THELLERSEN MARIANNE , PATKAR ANANT , PETERSEN AABY , ROYER C , KRETZSCHMAR TITUS
IPC分类号： C12N15/09 , C11D3/386 , C12N9/20

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