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    Method to isolate mutants and to clone the complementing gene
    6.
    发明授权
    Method to isolate mutants and to clone the complementing gene 失效
    分离突变体并克隆补体基因的方法

    公开(公告)号：US06177261B1

    公开(公告)日：2001-01-23

    申请号：US08981729

    申请日：1997-12-23

    申请人： Leendert Hendrik De Graaff ,  Henri{umlaut over (e)}tta Catharina Van Den Broeck ,  Jacob Visser

    发明人： Leendert Hendrik De Graaff ,  Henri{umlaut over (e)}tta Catharina Van Den Broeck ,  Jacob Visser

    IPC分类号： C12N115

    CPC分类号： C12N15/635 ,  C07K14/38 ,  C07K14/4705 ,  C12N9/0006 ,  C12N9/2434 ,  C12N9/248 ,  C12N9/88 ,  C12N15/1027 ,  C12N15/63 ,  C12N15/65 ,  C12N15/80 ,  C12Q1/02 ,  C12Y302/01003 ,  C12Y302/01008 ,  C12Y302/01015 ,  C12Y302/01037 ,  C12Y302/01055 ,  C12Y302/01099

    摘要： The subject invention lies in the field of microorganism mutation and selection of the mutants. In particular, the invention is directed at obtaining metabolic mutants in a simple, direct and specific manner. In a preferred embodiment it is also possible to obtain desired mutants not comprising recombinant DNA, thereby facilitating incorporation thereof in products for human consumption or application, due to shorter legislative procedures. The method according to the invention involves random mutation and specific selection of the desired metabolic mutant. A nucleic acid cassette comprising a nucleic acid sequence encoding a bidirectional marker, said nucleic acid cassette further comprising a basic transcriptional unit operatively linked to the nucleic acid sequence encoding the bidirectional marker and said nucleic acid cassette further comprising an inducible enhancer or activator sequence linked to the basic transcription unit in such a manner that upon induction of the enhancer or activator sequence the bidirectional marker encoding nucleic acid sequence is expressed, said inducible enhancer or activator sequence being driven from a gene associated with metabolism is claimed as is application thereof in a selection method for mutants. In addition a regular gene xlnR encoding an activating regulator of an inducible enhancer or activator sequence and application of said gene and/or its expression product in overexpression of homologous or heterologous protein or peptide is described. Knockout mutants wherein said gene is absent or inactivated and mutants with increased or decreased DNA binding capacity are also claimed.

    摘要翻译： 本发明在于微生物突变和突变体的选择领域。 特别地，本发明涉及以简单，直接和具体的方式获得代谢突变体。 在优选的实施方案中，由于较短的立法程序，也可以获得不包含重组DNA的所需突变体，从而便于将其并入用于人类消费或应用的产品中。 根据本发明的方法涉及随机突变和所需代谢突变体的特异性选择。 一种核酸盒，其包含编码双向标记的核酸序列，所述核酸盒还包含与编码双向标记的核酸序列可操作地连接的碱性转录单位，并且所述核酸盒还包含可诱导的增强子或活化剂序列 所述基本转录单元以这样一种方式使得在诱导增强子或活化剂序列时表达编码双向标记核酸序列的核酸序列，所述诱导型增强子或活化剂序列是从与代谢相关的基因驱动的，作为其选择 突变体方法。 此外，描述了编码诱导型增强子或激活物序列的活化调节物的正常基因xlnR以及所述基因和/或其表达产物在同源或异源蛋白质或肽的过表达中的应用。 其中所述基因不存在或失活的敲除突变体以及DNA结合能力增加或降低的突变体也被要求保护。