公开(公告)号：US07172882B2

公开(公告)日：2007-02-06

申请号：US10511327

申请日：2003-04-14

申请人： Harri Savilahti ,  Ville Tieaho

发明人： Harri Savilahti ,  Ville Tieaho

IPC分类号： C12P19/34 ,  C12N15/66

CPC分类号： C12N15/102 ,  C12N15/902

摘要： The present invention describes an in vitro transposition-based methodology for generation of deletion derivatives of polypeptides. An artificial transposon containing at least partly within its transposon ends a modification with translation stop codons in three reading frames is provided. In the method, transposition complexes are assembled using the modified transposon and essentially random integrations into the target plasmid, containing a polypeptide coding nucleic acid of interest, are recovered as a plasmid pool. Subsequent manipulation steps including restriction enzyme digestions and ligation result in pools of mutant clones from which deletion derivatives of a polypeptide coding nucleic acid of interest and its respective deletion polypeptides could be produced.

摘要翻译： 本发明描述了用于产生多肽的缺失衍生物的基于体外转座的方法。 提供至少部分含有其转座子的人工转座子在三个阅读框中结束具有翻译终止密码子的修饰。 在该方法中，使用修饰的转座子组装转座复合体，并将含有编码目标核酸的多肽的目标质粒的基本随机整合作为质粒库回收。 随后的包括限制酶消化和连接的操作步骤导致突变克隆的库，从中可以产生编码目的核酸的多肽的缺失衍生物及其各自的缺失多肽。

公开(公告)号：US20050208616A1

公开(公告)日：2005-09-22

申请号：US10511327

申请日：2003-04-14

申请人： Harri Savilahti ,  Ville Tieaho

发明人： Harri Savilahti ,  Ville Tieaho

IPC分类号： C12N15/10 ,  C12N15/90 ,  C12P21/06 ,  C12N15/74

CPC分类号： C12N15/102 ,  C12N15/902

摘要： The present invention describes an in vitro transposition-based methodology for generation of deletion derivatives of polypeptides. An artificial transposon containing at least partly within its transposon ends a modification with translation stop codons in three reading frames is provided. In the method, transposition complexes are assembled using the modified transposon and essentially random integrations into the target plasmid, containing a polypeptide coding nucleic acid of interest, are recovered as a plasmid pool. Subsequent manipulation steps including restriction enzyme digestions and ligation result in pools of mutant clones from which deletion derivatives of a polypeptide coding nucleic acid of interest and its respective deletion polypeptides could be produced.

摘要翻译： 本发明描述了用于产生多肽的缺失衍生物的基于体外转座的方法。 提供至少部分含有其转座子的人工转座子在三个阅读框中结束具有翻译终止密码子的修饰。 在该方法中，使用修饰的转座子组装转座复合体，并将含有编码目标核酸的多肽的目标质粒的基本随机整合作为质粒库回收。 随后的包括限制酶消化和连接的操作步骤导致突变克隆的库，从中可以产生编码目的核酸的多肽的缺失衍生物及其各自的缺失多肽。

公开(公告)号：US06344341B1

公开(公告)日：2002-02-05

申请号：US09446920

申请日：1999-12-30

申请人： Sirkka Keränen ,  Jaana Toikkanen ,  Ville Tieaho ,  Hans Söderlund

发明人： Sirkka Keränen ,  Jaana Toikkanen ,  Ville Tieaho ,  Hans Söderlund

IPC分类号： C12P2106

CPC分类号： C12N15/81 ,  C07K14/39

摘要： This invention relates to recombinant-DNA-technology. Specifically this invention relates to new recombinant yeast cells transformed with SEB1 gene. Yeast cells transformed with several copies of SEB1 gene, or overexpressing the Seb1 protein by some other means, have an increased capacity to produce secreted foreign or endogenous proteins. Further, said new recombinant cells, when transformed with genes expressing suitable hydrolytic enzymes can utilize appropriate macromolecular compounds more efficiently, which results in increased cell mass production and/or more versatile utilization of the compounds in relevant biotechnical applications.

摘要翻译： 本发明涉及重组DNA技术。 具体地说，本发明涉及用SEB1基因转化的新的重组酵母细胞。 用几个拷贝的SEB1基因转化的酵母细胞，或通过其他方法过表达Seb1蛋白，具有增加产生分泌的外源或内源蛋白质的能力。 此外，当用表达合适的水解酶的基因转化时，所述新的重组细胞可以更有效地利用合适的大分子化合物，这导致在相关生物技术应用中化合物增加的细胞大量产生和/或更通用的利用。