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    Method of heterologous cloning of gene in bacillus microorganism
    3.
    发明公开
    Method of heterologous cloning of gene in bacillus microorganism 失效
    Verfahren zur heterologen Klonierung eines Ein Gens in einem Bacillus-Mikroorganismus。

    公开(公告)号：EP0108301A2

    公开(公告)日：1984-05-16

    申请号：EP83110394.0

    申请日：1983-10-19

    申请人： SOLVAY ENZYMES, INC. (a Delaware corporation)

    发明人： Grosch, Josephine ,  Wilson, Gary A. ,  Wollweber, Karen ,  Yehle, Clifford O.

    IPC分类号： C12N15/56 ,  C12N15/57 ,  C12N15/75 ,  C12N9/28 ,  C12N9/34 ,  C12N9/54 ,  C12R1/07

    CPC分类号： C12N9/2417 ,  C12N9/2428 ,  C12N9/54 ,  C12N9/92 ,  C12N15/64 ,  C12N15/75 ,  C12Q1/40

    摘要： A method for heterologous cloning of a gene encoding α-amylase, protease, amyloglucosidase or glucose isomerase. The method involves the use of a bifunctional plasmid which can replicate in both E. coli and B. subtilis, and contains at least two selectable markers. A gene fragment for the enzyme is incorporated into the plasmid, leaving at least one functional selectable marker and a nonfunctional marker, producing a mixture of recombinant hybrid plasmids and the original bifunctional plasmid. This mixture is incorporated into E. coli microorganisms and transformants seiected. The transformants which contain the recombinant plasmid are identified, and plasmid DNA extracted. The plasmid DNA is used to transform Bacillus host microorganisms and transformants containing the recombinant plasmids are selected by means of the functional selectable marker.

    摘要翻译： 用于异源克隆编码α-淀粉酶，蛋白酶，淀粉葡糖苷酶或葡萄糖异构酶的基因的方法。 该方法涉及使用可在大肠杆菌和枯草芽孢杆菌中复制并含有至少两种可选择标记的双功能质粒。 将该酶的基因片段并入质粒中，留下至少一个功能选择标记和非功能标记，产生重组杂交质粒和原始双功能质粒的混合物。 将该混合物并入大肠杆菌微生物和转化体中。 鉴定含有重组质粒的转化体，提取质粒DNA。 质粒DNA用于转化芽孢杆菌宿主微生物，通过功能选择标记选择含有重组质粒的转化体。