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    Mutants of Deoxycytidine Kinase Having Extended Enzymatic Activity
    1.
    发明申请
    Mutants of Deoxycytidine Kinase Having Extended Enzymatic Activity 审中-公开
    具有延长酶活性的脱氧胞苷激酶突变体

    公开(公告)号：US20100136693A1

    公开(公告)日：2010-06-03

    申请号：US12463239

    申请日：2009-05-08

    申请人： Philippe Marliere ,  Sylvie Pochet ,  Madeleine Bouzon

    发明人： Philippe Marliere ,  Sylvie Pochet ,  Madeleine Bouzon

    IPC分类号： C12N15/01 ,  C12N9/12 ,  C12P19/38

    CPC分类号： C12N15/102 ,  C12N9/1205 ,  C12N15/01 ,  C12N15/1058

    摘要： The invention relates to a method for artificial in vivo evolution of proteins, said method making it possible to bring about the evolution of a protein X by complementation of a relative protein Y, X and Y both belonging to the same class of enzyme commission (EC) nomenclature or belonging to related classes. The mutants D133E and R104Q of desoxycytidine kinase (DCK) were obtained; both of said mutations result in acquisition of thymidine kinase activity by DCK.

    摘要翻译： 本发明涉及蛋白质人体体内进化的方法，所述方法使得可以通过相互作用的相关蛋白Y，X和Y的互补来实现蛋白X的进化，这两者属于同一类酶委员会（EC ）命名或属于相关类。 获得脱氧胞苷激酶（DCK）的突变体D133E和R104Q; 所述两个突变导致通过DCK获得胸苷激酶活性。

    Method for the in vivo modification of the synthesis activity of a metabolite by means of the modification of a gene the activity of which is not the original activity
    5.
    发明授权
    Method for the in vivo modification of the synthesis activity of a metabolite by means of the modification of a gene the activity of which is not the original activity 有权
    通过改变基因的体内修饰代谢物的合成活性的方法，其活性不是原始活性

    公开(公告)号：US07892838B2

    公开(公告)日：2011-02-22

    申请号：US11735148

    申请日：2007-04-13

    申请人： Pierre-Alexandre Kaminski ,  Philippe Marliere

    发明人： Pierre-Alexandre Kaminski ,  Philippe Marliere

    IPC分类号： C12P21/06 ,  C12P19/30 ,  C12N9/10 ,  C12N1/20 ,  C12N15/00 ,  C12N15/74 ,  C07H21/02 ,  C07H21/04

    CPC分类号： C12N15/1058 ,  C12N15/102

    摘要： The invention relates to a method for altering a protein X such as to modify the characteristics thereof by a) obtaining the mutants X* of the sequence coding for protein X, by means of aleatory mutagenesis, b) transformation of cells with a phenotype [P-] with vectors comprising the mutated nucleic acids obtained in step (a) which code for proteins X*, where P-signifies that said cells are auxotrophic for substance P, P begin the product of the action of X on the natural substrate thereof S, c) culturing said cells in a medium comprising a substrate S*, S* being an analogue of the natural substrate S of the protein X, d) selection of the cells [P-:: X*] which have survived step c) in which the proteins X* can biosynthesise the product P from the substrate S*. The invention further relates to mutated proteins X, nucleic acids, expression vectors, host cells comprising a vector, use of N-dideoxyribosyl transferases for the transfer of a dideoxyribose (ddR) from a dideoxyribonucleoside to another nucleoside, a method for production of compounds comprising a step using a mutated protein and a strain of E. coli.

    摘要翻译： 本发明涉及一种用于改变蛋白质X的方法，例如通过以下方式改变其特征：a）通过亲和诱变获得编码蛋白质X的序列的突变体X *，b）用表型转化的细胞[P - ]，其中包含步骤（a）中获得的编码蛋白质X *的突变核酸的载体，其中P表示所述细胞对于物质P是营养缺陷型的，P开始在其天然底物上的作用X的产物S c）在包含蛋白质X的天然底物S的类似物的底物S *，S *的培养基中培养所述细胞，d）选择在步骤c）中存活的细胞[P- :: X *]， 其中蛋白质X *可以从底物S *生物合成产物P. 本发明进一步涉及突变蛋白X，核酸，表达载体，包含载体的宿主细胞，使用N-二脱氧核糖基转移酶将二脱氧核糖（ddR）从双脱氧核糖核苷转移至另一核苷的方法，包括 使用突变蛋白和大肠杆菌菌株的步骤。

    Method and device for selecting accelerated proliferation of living cells in suspension
    8.
    发明授权
    Method and device for selecting accelerated proliferation of living cells in suspension 有权
    选择悬浮液中活细胞加速增殖的方法和装置

    公开(公告)号：US06686194B1

    公开(公告)日：2004-02-03

    申请号：US09857226

    申请日：2001-06-01

    申请人： Rupert Mutzel ,  Philippe Marliere

    发明人： Rupert Mutzel ,  Philippe Marliere

    IPC分类号： C12M136

    CPC分类号： C12M23/58 ,  C12M37/00 ,  Y10S435/813

    摘要： The present invention relates to a method and a device for selecting accelerated proliferation of living cells in suspension. The culture apparatus (2) of the present invention enables cells to proliferate in suspension over unlimited periods of time. Natural selection results in the accumulation of genetic variants which are increasingly better adapted to the chosen culture conditions. The organisms used can be prokaryotic or eukaryotic. The organisms used can be naturally occurring organisms or genetically modified organisms. The culture apparatus of the present invention is also suitable for using continuous, periodical or conditional culture conditions. The physical and chemical characteristics of the culture media used can be chosen by the user. The requirement that a population of cells proliferates exclusively in suspension in continuous culture conditions is satisfied by the periodical transfer of the organism suspension from a first culture vessel into a second culture vessel. After the transfer, the first culture vessel is subjected to a sterilizing treatment and the sterilizing agent is optionally neutralized, so that the first culture vessel is ready for the culture to be transferred back from the second culture vessel. The second culture vessel is then sterilized and neutralized.

    摘要翻译： 本发明涉及用于选择悬浮液中活细胞加速增殖的方法和装置。 本发明的培养装置（2）能够使细胞在无限期内悬浮增殖。 自然选择导致遗传变异体的积累，其越来越适应于所选择的培养条件。 所用的生物体可以是原核生物或真核生物。 所使用的生物体可以是天然存在的生物体或转基因生物。 本发明的培养装置也适用于连续，周期或条件培养条件。 使用的培养基的物理和化学特性可由用户选择。 通过将生物体悬浮液从第一培养容器定期转移到第二培养容器中，满足在连续培养条件下悬浮的细胞群体增殖的要求。 转移后，对第一培养容器进行灭菌处理，任选地中和杀菌剂，使第一培养容器准备从第二培养容器中转移回培养物。 然后将第二培养容器消毒并中和。

    Method for obtaining new life forms
    9.
    发明授权
    Method for obtaining new life forms 有权
    获得新生命形式的方法

    公开(公告)号：US07960103B2

    公开(公告)日：2011-06-14

    申请号：US11519978

    申请日：2006-09-13

    申请人： Rupert Mutzel ,  Philippe Marliere ,  Didier Mazel

    发明人： Rupert Mutzel ,  Philippe Marliere ,  Didier Mazel

    IPC分类号： C12Q1/68

    CPC分类号： C12R1/19 ,  C12N9/1014 ,  C12N9/80 ,  C12N15/1058 ,  C12N15/70 ,  C12P21/02 ,  C12Y305/01088 ,  C40B10/00

    摘要： The present invention relates to a method for generating a novel form of life comprising the steps consisting of: a) irreversible alteration of the genome of a microbial clone; b) cultivation of a vast population of microbial cells originating from the altered clone obtained in step a) during numerous generations under conditions allowing selection for a higher and stable proliferation rate; c) isolation of descendant clones within the cultivated population of step b) still bearing the alteration of step a).

    摘要翻译： 本发明涉及一种产生新生命形式的方法，包括以下步骤：a）微生物克隆的基因组的不可逆改变; b）在允许选择更高和稳定的增殖速率的条件下，在许多代中培养源自步骤a）中获得的改变的克隆的大量微生物细胞群; c）分离步骤b）的栽培种群中的后代克隆仍然具有步骤a）的改变。

    Cells and method for producing proteins comprising an unconventional amino acid
    10.
    发明授权
    Cells and method for producing proteins comprising an unconventional amino acid 失效
    用于产生包含非常规氨基酸的蛋白质的细胞和方法

    公开(公告)号：US07736899B1

    公开(公告)日：2010-06-15

    申请号：US09830669

    申请日：1999-10-28

    申请人： Philippe Marliere ,  Volker Doring ,  Henning Mootz

    发明人： Philippe Marliere ,  Volker Doring ,  Henning Mootz

    IPC分类号： C12N15/00 ,  C12N1/20 ,  C12P21/00

    CPC分类号： C12N15/1058 ,  A61K38/00 ,  A61K48/00 ,  C12N15/67 ,  C12P21/02

    摘要： The present invention relates to a method for providing bacterial or yeast cells with the capacity to produce a protein, the amino acid sequence of which comprises at least one unconventional amino acid. The method involves (a) introducing at least one missense mutation in a target codon of a gene encoding a protein required for the growth of the bacterial or yeast cells, where the mutated protein synthesized from the mutated gene is not functional in the bacterial or yeast cells. The method also involves (b) selecting the bacterial or yeast cells obtained in (a) in a culture medium which (1) does not contain a nutrient compensating for the loss of functionality of the mutated protein and (2) contains an unconventional amino acid which restores the functionality of the protein required for growth of the bacterial or yeast cells, in which the unconventional amino acid is that encoded by the target codon. The method also involves culturing the bacterial or yeast cells obtained in (b) in a culture medium containing the amino acid encoded by the target codon.

    摘要翻译： 本发明涉及提供具有产生蛋白质的能力的细菌或酵母细胞的方法，所述蛋白质的氨基酸序列包含至少一种非常规氨基酸。 该方法包括（a）在编码细菌或酵母细胞生长所需的蛋白质的基因的靶标密码子中引入至少一个错义突变，其中从突变基因合成的突变蛋白在细菌或酵母中不起作用 细胞。 该方法还包括（b）在培养基中选择（a）中获得的细菌或酵母细胞，其中（1）不含营养物，补偿突变蛋白的功能丧失，（2）含有非常规氨基酸 其恢复细菌或酵母细胞生长所需的蛋白质的功能，其中非常规氨基酸是由靶标密码子编码的。 该方法还包括在含有由目标密码子编码的氨基酸的培养基中培养（b）中获得的细菌或酵母细胞。