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    • 2. 发明授权
    • Methods for generating recombined polynucleotides
    • 产生重组多核苷酸的方法
    • US06541207B1
    • 2003-04-01
    • US09687301
    • 2000-10-13
    • Jesper VindTorben Vedel Borchert
    • Jesper VindTorben Vedel Borchert
    • C12N1500
    • C12N15/1093C12N15/1027
    • A method for in vitro construction of a library of recombined homologous polynucleotides from a number of different starting DNA templates and primers by induced template shifts during an polynucleotide synthesis is described, whereby A. extended primers are synthesized by a) denaturing the DNA templates b) annealing primers to the templates, c) extending the said primers by use of a polymerase, d) stop the synthesis, and e) separate the extended primers from the templates, B. a template shift is induced by a) isolating the extended primers from the templates and repeating steps A.b) to A.e) using the extended primers as both primers and templates, or b) repeating steps A.b) to A.e), C. this process is terminated after an appropriate number of cycles of process steps A. and B.a), A. and B.b), or combinations thereof. Optionally the polynucleotides are amplified in a standard PCR reaction with specific primers to selectively amplify homologous polynucleotides of interest.
    • 描述了在多核苷酸合成期间通过诱导的模板位移从许多不同的起始DNA模板和引物体外构建重组的同源多核苷酸文库的方法,其中A. 通过a)使DNA模板变性b)将引物退火到模板来合成延伸的引物,c)使用聚合酶扩增所述引物,d)停止合成,并且将扩展的引物与模板B分开。 通过以下步骤诱导模板位移:a)使扩展引物与模板分离,并重复步骤A.b)至A.e),使用扩展引物作为引物和模板,orb)重复步骤A.b)至A.e),C。 该方法在方法步骤A和B.a),A.和B.b)或其组合的适当数目的循环后终止。可选地,在与特异性引物的标准PCR反应中扩增多核苷酸以选择性扩增目的同源多核苷酸。
    • 5. 发明授权
    • Alpha-amylase mutants
    • α-淀粉酶突变体
    • US08323946B2
    • 2012-12-04
    • US13161931
    • 2011-06-16
    • Allan SvendsenHenrik Bisgard-FrantzenTorben Vedel Borchert
    • Allan SvendsenHenrik Bisgard-FrantzenTorben Vedel Borchert
    • C12N9/00C12N9/30C12N1/20C12N15/00C07H21/04
    • C11D3/386C12N9/2417
    • The present invention relates to a method of constructing a variant of a parent Termamyl-like alpha-amylase, which variant has alpha-amylase activity and at least one altered property as compared to the parent alpha-amylase, comprises i) analyzing the structure of the parent Termamyl-like alpha-amylase to identify at least one amino acid residue or at least one structural part of the Termamyl-like alpha-amylase structure, which amino acid residue or structural part is believed to be of relevance for altering the property of the parent Termamyl-like alpha-amylase (as evaluated on the basis of structural or functional considerations), ii) constructing a Termamyl-like alpha-amylase variant, which as compared to the parent Termamyl-like alpha-amylase, has been modified in the amino acid residue or structural part identified in i) so as to alter the property, and, optionally, iii) testing the resulting Termamyl-like alpha-amylase variant with respect to the property in question.
    • 本发明涉及一种构建母体类戊酰样α-淀粉酶的变体的方法,该变体具有与母体α-淀粉酶相比具有α-淀粉酶活性和至少一种改变的性质,包括:i)分析 鉴定至少一个氨基酸残基或类甲酰样α-淀粉酶结构的至少一个结构部分,所述氨基酸残基或结构部分被认为与改变其性质有关的相关性 母体类戊酰样α-淀粉酶(根据结构或功能考虑进行评估),ii)构建与母体类戊酰样α-淀粉酶相比的类戊酰样α-淀粉酶变体,已经在 在i)中鉴定的氨基酸残基或结构部分,以改变其性质,以及任选地,iii)相对于所讨论的性质测试所得的类戊酰样α-淀粉酶变体。