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    Genome sequence based targeted cloning of DNA fragments
    26.
    发明公开
    Genome sequence based targeted cloning of DNA fragments 审中-公开
    Gezieltes Klonen von DNA-Fragmenten auf Genomsequenzbasis

    公开(公告)号：EP2677034A1

    公开(公告)日：2013-12-25

    申请号：EP12172389.4

    申请日：2012-06-18

    申请人： LEK Pharmaceuticals d.d.

    发明人： The designation of the inventor has not yet been filed

    IPC分类号： C12N15/70

    CPC分类号： C12N15/70

    摘要： The present invention belongs to the field of molecular biology, microbiology and microbial genetics and specifies the plasmid rescue method for targeted cloning of DNA fragments from a donor organism into a host cell, comprising inter alia the steps of transferring a Bacterial Artificial Chromosome (BAC)-vector that comprises a site-specific recombination system containming an integrase and an attP site into a donor organism, wherein said donor organism does not contain a functional attB site. Further, the present invention refers to a plasmid rescue method for targeted cloning of DNA fragments from a donor organism into a host cell, comprising inter alia the steps of transferring a BAC-vector that comprises a site-specific recombination system containing an attP site however no functional integrase into a donor organism, wherein in a further step a functional integrase has to be introduced into a final host cell. Further, the present invention refers to a vector comprising inter alia a site-specific recombination system containing an attP site however no functional integrase, and to a vector comprising inter alia a site-specific recombination system containing an integrase and an attP site and a DNA sequence suitable for homologous recombination. The present invention further refers to the use of said vectors for cloning and/or transferring of a DNA fragment. Additionally, the present invention refers to suitable donor organisms comprising the BAC vectors according to the present invention, to an expression system comprising a host cell and a BAC vector, as well as to the use of said expression system for cloning and/or transfer of the DNA fragment, and /or for expression of said DNA fragment.

    摘要翻译： 本发明属于分子生物学，微生物学和微生物遗传学领域，并且具体规定了将供体生物体的DNA片段靶向克隆到宿主细胞中的质粒拯救方法，其特别包括将细菌人工染色体（BAC） - 载体，其包含将供体生物体的整合酶和attP位点包含的位点特异性重组系统，其中所述供体生物体不含有功能性attB位点。 此外，本发明涉及用于将来自供体生物体的DNA片段靶向克隆到宿主细胞中的质粒拯救方法，其特别包括转移包含含有attP位点的位点特异性重组系统的BAC载体的步骤 没有功能整合酶进入供体生物体，其中在另外的步骤中，必须将功能性整合酶引入最终宿主细胞。 此外，本发明涉及包含尤其是含有attP位点但不包括功能整合酶的位点特异性重组系统的载体，以及包含尤其是含有整合酶和attP位点的位点特异性重组系统和DNA 适合同源重组的序列。 本发明还涉及所述载体用于克隆和/或转移DNA片段的用途。 另外，本发明涉及包含本发明的BAC载体的合适的供体生物体，包括宿主细胞和BAC载体的表达系统，以及使用所述表达系统克隆和/或转移 DNA片段和/或用于表达所述DNA片段。

    ENZYMATIC SYNTHESIS OF ACTIVE PHARMACEUTICAL INGREDIENT AND INTERMEDIATES THEREOF
    27.
    发明公开
    ENZYMATIC SYNTHESIS OF ACTIVE PHARMACEUTICAL INGREDIENT AND INTERMEDIATES THEREOF 有权
    ENZYMATISCHE SYNTHESE EINES PHARMAZEUTISCHEN WIRKSTOFFS UND ZWISCHENPRODUKTE DAVON

    公开(公告)号：EP2655650A2

    公开(公告)日：2013-10-30

    申请号：EP11811536.9

    申请日：2011-12-20

    申请人： LEK Pharmaceuticals d.d.

    发明人： MRAK, Peter ,  ZOHAR, Tadeja ,  OSLAJ, Matej ,  KOPITAR, Gregor

    IPC分类号： C12P17/06 ,  C12P17/16 ,  C12N9/88 ,  C12N15/70 ,  C12N9/00

    CPC分类号： C12P17/06 ,  C12N9/0006 ,  C12N9/88 ,  C12N15/70 ,  C12P17/10 ,  C12P17/12 ,  C12P17/16

    摘要： The present invention discloses a process for preparing an active pharmaceutical ingredient (API) or intermediates thereof, notably particular step in the synthesis of an intermediate useful for example in the preparation of statins, by using an enzyme capable of catalyzing oxidation or dehydrogenation. The invention further provides an expression system effectively translating said enzyme. In addition, the invention relates to a specific use of such enzyme for preparing API or intermediate thereof, and in particular for preparing statin or intermediate thereof.

    摘要翻译： 本发明公开了一种制备活性药物成分（API）或其中间体的方法，特别是通过使用能够催化氧化或脱氢的酶合成可用于制备他汀类的中间体的特别步骤。 本发明还提供了有效翻译所述酶的表达系统。 此外，本发明涉及这种酶用于制备API或其中间体，特别是用于制备他汀类或其中间体的具体用途。