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1. 发明授权

US5837509A Recombinant lactic acid bacterium containing an inserted promoter and method of constructing same 失效
标题翻译：含有插入启动子的重组乳酸菌及其构建方法
公开(公告)号：US5837509A
公开(公告)日：1998-11-17
申请号：US179557
申请日：1995-03-01
申请人： Hans Israelsen , Egon Bech Hansen , Eric Johansen , Soeren Michael Madsen , Dan Nilsson , Astrid Vrang
发明人： Hans Israelsen , Egon Bech Hansen , Eric Johansen , Soeren Michael Madsen , Dan Nilsson , Astrid Vrang
IPC分类号： C12N1/21 , C12N15/74 , C12N15/77 , C12N15/11 , C12P21/02
CPC分类号： C12N15/746 , C12N15/77 , Y10S435/853
摘要： A method of isolating a lactic acid bacterial DNA fragment comprising a promoter, the method comprising introducing a DNA molecule comprising a transposable element comprising a promoterless structural gene as a promoter probe gene into a population of a lactic acid bacterium, methods of constructing a recombinant lactic acid bacterium comprising a regulatable promoter by using the above method, a recombinant lactic acid bacterium comprising a gene coding for a desired gene product and operably linked thereto a regulatable lactic acid bacterial promoter not natively associated with the gene, the use of such a recombinant lactic acid bacterium and recombinant plasmids comprising a regulatable lactic acid bacterial promoter.
摘要翻译： PCT No.PCT / DK94 / 00004 Sec。 371 1995年3月1日 102（e）1995年3月1日PCT 1994年3月1日PCT PCT。 WO94 / 16086 PCT出版物日期：1994年7月21日一种分离含有启动子的乳酸菌DNA片段的方法，该方法包括将包含无启动子结构基因的转座元件作为启动子探针基因的DNA分子引入乳酸菌群中，通过使用上述方法构建包含可调节启动子的重组乳酸菌，重组乳酸菌，其包含编码所需基因产物的基因，并与其可操作地连接与该基因无天然关联的可调节乳酸细菌启动子，使用的这种重组乳酸菌和包含可调节的乳酸细菌启动子的重组质粒。

2. 发明授权

US07358067B2 Fermentation method for production of heterologous gene products in lactic acid bacteria 有权
标题翻译：用于在乳酸菌中生产异源基因产物的发酵方法
公开(公告)号：US07358067B2
公开(公告)日：2008-04-15
申请号：US09982531
申请日：2001-10-19
申请人： Astrid Vrang , Søren Michael Madsen , Lars Bredmose , Peter Ravn , Jose Arnau , Mads Gronvald Johnsen , Anne Cathrine Steenberg , Hans Israelsen
发明人： Astrid Vrang , Søren Michael Madsen , Lars Bredmose , Peter Ravn , Jose Arnau , Mads Gronvald Johnsen , Anne Cathrine Steenberg , Hans Israelsen
IPC分类号： C12P21/00 , C12N1/21
CPC分类号： C12N15/74 , C12N1/00 , C12N9/22 , C12N15/67
摘要： The invention provides a fed-batch or continuous process for producing heterologous peptides, polypeptides or proteins in lactic acid bacteria, comprising the steps of (i) constructing a recombinant lactic acid bacterium comprising a nucleotide sequence coding for the heterologous peptide, polypeptide or protein, and appropriate regulatory nucleotide sequences such as regulatable promoters and signal peptides to control the expression of the coding sequence and the secretion of gene product, (ii) cultivating the recombinant bacterium under or continuous cultivation conditions to express the gene, and (iii) harvesting the recombinant bacterium or the gene product. Preferably, the cultivation medium is a chemically defined or synthetic medium optionally supplemented with yeast extract.
摘要翻译：本发明提供了用于在乳酸菌中产生异源肽，多肽或蛋白质的补料分批或连续方法，包括以下步骤：（i）构建重组乳酸菌，其包含编码异源肽，多肽或蛋白质的核苷酸序列，以及适当的调控核苷酸序列如可调节的启动子和信号肽，以控制编码序列的表达和基因产物的分泌，（ii）在培养条件下或连续培养条件下培养重组细菌以表达基因，和（iii）收获重组细菌或基因产物。优选地，培养基是任选地补充有酵母提取物的化学上定义的或合成的培养基。

3. 发明授权

US07186815B2 Method of isolating secretion signals in lactic acid bacteria and novel secretion signals isolated from Lactococcus lactis 有权
标题翻译：分离乳酸菌分泌信号的方法和从乳酸乳球菌分离的新型分泌信号
公开(公告)号：US07186815B2
公开(公告)日：2007-03-06
申请号：US10431577
申请日：2003-05-08
申请人： Peter Ravn , Soeren Michael Madsen , Astrid Vrang , Hans Israelsen , Mads Groenvold Johnsen , Lars Bredmose , José Arnau
发明人： Peter Ravn , Soeren Michael Madsen , Astrid Vrang , Hans Israelsen , Mads Groenvold Johnsen , Lars Bredmose , José Arnau
IPC分类号： C07H21/02 , C07H21/04 , C07N7/00 , C07N15/00 , C07N15/09
CPC分类号： C12N15/1034 , C07K2319/02 , C07K2319/036 , C07K2319/61 , C12N15/625 , C12N15/746
摘要： A method of identifying nucleotide sequences coding for signal peptides in lactic acid bacteria, using a DNA molecule comprising a transposon including a promoterless reporter gene from which DNA molecule a region between the LR and the reporter gene is deleted and the DNA molecule comprises a DNA sequence coding for a secretion reporter molecule. By deleting the region between the LR and the reporter gene, stop codons in-frame with the secretion reporter molecule is removed which upon transposition permits translational fusions from upstream the LR.
摘要翻译：使用包含转座子的DNA分子的DNA分子的方法，所述DNA分子包含转座子，所述转座子包含DNA分子，LR分子和报告基因之间的区域由DNA启动子报告基因组成，DNA分子包含DNA序列编码分泌报道分子。通过删除LR和报告基因之间的区域，消除了与分泌报告分子的框内的密码子，其在转座后允许从LR的上游进行翻译融合。

4. 发明授权

US06133023A Lactic acid bacterial regulatable expression system 失效
标题翻译：乳酸菌可调节表达系统
公开(公告)号：US06133023A
公开(公告)日：2000-10-17
申请号：US981601
申请日：1997-12-29
申请人： Soeren Michael Madsen , Astrid Vrang , Jose Arnau , Peter Ravn , Mads Groenvald Johnsen , Hans Israelsen
发明人： Soeren Michael Madsen , Astrid Vrang , Jose Arnau , Peter Ravn , Mads Groenvald Johnsen , Hans Israelsen
IPC分类号： A23L29/00 , C07K14/35 , C12N1/21 , C12N15/31 , C12N15/74 , C12N15/00 , C12N1/20 , C12N7/00 , C12P19/34
CPC分类号： C12N15/746 , C07K14/35
摘要： Expression vectors capable of being replicated in lactic acid bacterial cells comprising a promoter region comprising (a) a promoter sequence element the function of which is regulatable by an environmental or growth condition factor and (b) at least one further nucleotide sequence element, the position, orientation, presence and/or sequence of which element has a regulatory effect on the expression of a gene operably linked to the promoter region in which vectors the position, orientation, presence and/or sequence of at least one of said elements (a) or (b) is modified relative to the position, orientation, presence and/or sequence of the corresponding non-modified element whereby the expression of the gene is altered, and a lactic acid bacterium which is transformed with such a vector as defined above are provided. The recombinant cells containing such a regulatable or inducible gene expression system are useful as food or feed starter cultures or as strains for the production of gene products such as pharmaceutically or immunologically active compounds including oligo- or polypeptides derived from a Mycobacterium species including M. tuberculosis.
摘要翻译： PCT No.PCT / DK97 / 00341 Sec。 371日期1997年12月29日第 102（e）日期1997年12月29日PCT 1997年8月22日PCT公布。第WO98 / 10079号公报日期1998年3月12日能够复制在包含启动子区域的乳酸菌细胞中的表达载体包括（a）启动子序列元件，其功能由环境或生长条件因子调节，和（b）至少一个其它核苷酸序列元件，哪个元件的位置，取向，存在和/或序列对可操作地连接到启动子区的基因的表达具有调节作用，其中载体中的至少一个的位置，取向，存在和/或序列所述元件（a）或（b）相对于相应的未修饰元件的位置，取向，存在和/或序列被修饰，从而基因的表达被改变，并且用这种提供了如上所述的载体。含有这种可调节或可诱导基因表达系统的重组细胞可用作食物或饲料起始培养物或用作产生基因产物的菌株，例如药学或免疫活性化合物，包括衍生自包括结核分枝杆菌在内的分枝杆菌物种的寡肽或多肽。

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