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1.发明授权Cloning and expression of autogenes encoding RNA poly,erases of T7-like bacteriophages 失效编码RNA聚合酶的自身基因的克隆和表达，T7样噬菌体的消除
公开(公告)号：US5824528A
公开(公告)日：1998-10-20
申请号：US292081
申请日：1994-08-15
CPC分类号： C12N9/1247
摘要： This invention relates to the cloning and expression of autogenes encoding RNA polymerases of T7 and T7-like bacteriophages, in which the RNA polymerase gene is transcribed from a promoter which is recognized by the encoded RNA polymerase. Cloning of T7 autogenes was achieved by reducing the activity of the RNA polymerase sufficiently to permit host cell growth. T7 RNA polymerase activity was controlled by combining two independent methods: lac-repression of the recombinant lac operator-T7 promoter in the autogene and inhibition of the polymerase by T7 lysozyme. Expression systems for producing the RNA polymerases of T7 and other T7-like bacteriophages, and expression systems for producing selected gene products are described, as well as other related materials and methods.
摘要翻译： 本发明涉及编码T7和T7样噬菌体的RNA聚合酶的自身基因的克隆和表达，其中RNA聚合酶基因从编码的RNA聚合酶识别的启动子转录。 T7自身基因的克隆通过充分降低RNA聚合酶的活性以允许宿主细胞生长来实现。 T7 RNA聚合酶活性通过组合两种独立方法进行控制：在自身基因中重组lac操纵子-T7启动子的lac抑制和T7溶菌酶抑制聚合酶。 描述了用于产生T7和其他T7样噬菌体的RNA聚合酶的表达系统，以及用于产生所选基因产物的表达系统，以及其它相关材料和方法。
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2.发明授权公开(公告)号：US08399217B2
公开(公告)日：2013-03-19
申请号：US11741282
申请日：2007-04-27
申请人： F. William Studier
发明人： F. William Studier
摘要： A method for promoting and suppressing auto-induction of transcription of a cloned gene 1 of bacteriophage T7 in cultures of bacterial cells grown batchwise is disclosed. The transcription is under the control of a promoter whose activity can be induced by an exogenous inducer whose ability to induce said promoter is dependent on the metabolic state of said bacterial cells.
摘要翻译： 公开了一种在间歇生长的细菌细胞培养物中促进和抑制噬菌体T7的克隆基因1的转录自动诱导的方法。 转录受启动子的控制，其启动子的活性可由诱导所述启动子的能力依赖于所述细菌细胞的代谢状态的外源性诱导物诱导。
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公开(公告)号：US5790727A
公开(公告)日：1998-08-04
申请号：US796097
申请日：1997-02-05
CPC分类号： G02B6/26 , G02B6/3636 , G02B6/3652 , G02B6/3696 , Y10S385/901
摘要： A system and method are disclosed for efficient laser illumination of the interiors of multiple capillaries simultaneously, and collection of light emitted from them. Capillaries in a parallel array can form an optical waveguide wherein refraction at the cylindrical surfaces confines side-on illuminating light to the core of each successive capillary in the array. Methods are provided for determining conditions where capillaries will form a waveguide and for assessing and minimizing losses due to reflection. Light can be delivered to the arrayed capillaries through an integrated fiber optic transmitter or through a pair of such transmitters aligned coaxially at opposite sides of the array. Light emitted from materials within the capillaries can be carried to a detection system through optical fibers, each of which collects light from a single capillary, with little cross talk between the capillaries. The collection ends of the optical fibers can be in a parallel array with the same spacing as the capillary array, so that the collection fibers can all be aligned to the capillaries simultaneously. Applicability includes improving the efficiency of many analytical methods that use capillaries, including particularly high-throughput DNA sequencing and diagnostic methods based on capillary electrophoresis.
摘要翻译： 公开了一种系统和方法，用于同时对多个毛细管的内部进行有效的激光照射，以及从它们发射的光的收集。 平行阵列中的毛细管可以形成光波导，其中在圆柱形表面处的折射将侧向照射光限制在阵列中的每个连续毛细管的芯上。 提供了用于确定毛细管形成波导并用于评估和最小化反射损失的条件的方法。 光可以通过集成的光纤发射器或通过一对同轴对准的排列在阵列的相对侧上的阵列毛细管。 从毛细管内的材料发射的光可以通过光纤传送到检测系统，每个光纤从单个毛细管收集光，毛细管之间几乎没有串扰。 光纤的收集端可以是与毛细管阵列具有相同间隔的平行阵列，使得收集纤维都可以同时与毛细管对准。 适用性包括提高使用毛细血管的许多分析方法的效率，包括特别是基于毛细管电泳的高通量DNA测序和诊断方法。
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公开(公告)号：US20080187986A1
公开(公告)日：2008-08-07
申请号：US12062575
申请日：2008-04-04
申请人： F. William Studier
发明人： F. William Studier
IPC分类号： C12N1/20
摘要： A bacterial growth medium for promoting auto-induction of transcription of cloned DNA in cultures of bacterial cells grown batchwise is disclosed. The transcription is under the control of a lac repressor. Also disclosed is a bacterial growth medium for improving the production of a selenomethionine-containing protein or polypeptide in a bacterial cell, the protein or polypeptide being produced by recombinant DNA techniques from a lac or T7lac promoter, the bacterial cell encoding a vitamin B12-dependent homocysteine methylase. Finally, disclosed is a bacterial growth medium for suppressing auto-induction of expression in cultures of bacterial cells grown batchwise, said transcription being under the control of lac repressor.
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5.发明申请公开(公告)号：US20080187985A1
公开(公告)日：2008-08-07
申请号：US12062565
申请日：2008-04-04
申请人： F. William Studier
发明人： F. William Studier
IPC分类号： C12N1/20
摘要： A bacterial growth medium for promoting auto-induction of transcription of cloned DNA in cultures of bacterial cells grown batchwise is disclosed. The transcription is under the control of a lac repressor. Also disclosed is a bacterial growth medium for improving the production of a selenomethionine-containing protein or polypeptide in a bacterial cell, the protein or polypeptide being produced by recombinant DNA techniques from a lac or T7lac promoter, the bacterial cell encoding a vitamin B12-dependent homocysteine methylase. Finally, disclosed is a bacterial growth medium for suppressing auto-induction of expression in cultures of bacterial cells grown batchwise, said transcription being under the control of lac repressor.
摘要翻译： 公开了一种细菌生长培养基，其用于促进分批生长的细菌细胞培养物中克隆DNA的转录自动诱导。 转录受lac阻遏物的控制。 还公开了一种细菌生长培养基，用于改善细菌细胞中含有含硒蛋氨酸的蛋白质或多肽的产生，所述蛋白质或多肽通过来自lac或T7lac启动子的重组DNA技术产生，编码维生素B12依赖性的细菌细胞 高半胱氨酸甲基化酶。 最后，公开了一种细菌生长培养基，其用于抑制细菌生长在分批培养物中的表达的自身诱导，所述转录在lac阻抑物的控制下。
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6.发明授权
公开(公告)号：US5693489A
公开(公告)日：1997-12-02
申请号：US259560
申请日：1994-06-14
申请人： F. William Studier , Parichehre Davanloo , Alan H. Rosenberg , Barbara A. Moffatt , John J. Dunn
发明人： F. William Studier , Parichehre Davanloo , Alan H. Rosenberg , Barbara A. Moffatt , John J. Dunn
CPC分类号： C12N9/1247 , C12N15/70 , C12N15/73
摘要： This application describes a means to clone a functional gene for bacteriophage T7 RNA polymerase. Active T7 RNA polymerase is produced from the cloned gene, and a plasmid has been constructed that can produce the active enzyme in large amounts. T7 RNA polymerase transcribes DNA very efficiently and is highly selective for a relatively long promoter sequence. This enzyme is useful for synthesizing large amounts of RNA in vivo or in vitro, and is capable of producing a single RNA selectively from a complex mixture of DNAs. The procedure used to obtain a clone of the R7 RNA polymerase gene can be applied to other T7-like phages to obtain clones that produce RNA polymerases having different promoter specificities, different bacterial hosts, or other desirable properties. T7 RNA polymerase is also used in a system for selective, high-level synthesis of RNAs and proteins in suitable host cells.
摘要翻译： 本申请描述了克隆噬菌体T7RNA聚合酶的功能基因的方法。 活性T7 RNA聚合酶是从克隆的基因中产生的，已经构建出能大量产生活性酶的质粒。 T7 RNA聚合酶非常有效地转录DNA，对于较长的启动子序列具有高选择性。 该酶可用于在体内或体外合成大量的RNA，并且能够从DNA的复杂混合物中选择性地产生单一RNA。 用于获得R7 RNA聚合酶基因克隆的程序可以应用于其他T7样噬菌体，以获得产生具有不同启动子特异性，不同细菌宿主或其它所需性质的RNA聚合酶的克隆。 T7 RNA聚合酶还用于在合适的宿主细胞中选择性，高水平合成RNA和蛋白质的系统。
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公开(公告)号：US5550035A
公开(公告)日：1996-08-27
申请号：US187119
申请日：1994-01-26
CPC分类号： C12N9/1247 , C12N15/10 , C12N15/85
摘要： A transient expression system is disclosed that utilizes bacteriophage RNA polymerase in the presence of a DNA-based cytoplasmic virus to facilitate expression of a foreign gene in the cytoplasm of a eukaryotic cell.A method of expressing a foreign gene in the cytoplasm of a eukaryotic cell is also disclosed which comprises incorporating into the cytoplasm a DNA-based cytoplasmic virus, a suitable carrier comprising a gene for an RNA polymerase which gene is foreign to the carrier and to the cells, and a suitable carrier comprising a functional, cistron including a foreign gene flanked by a promotor sequence which is recognized by the RNA polymerase.
摘要翻译： 公开了一种瞬时表达系统，其在存在基于DNA的细胞质病毒的情况下利用噬菌体RNA聚合酶促进外源基因在真核细胞的细胞质中的表达。 还公开了在真核细胞的细胞质中表达外源基因的方法，其包括向细胞质中加入基于DNA的细胞质病毒，合适的载体，其包含用于载体外源的RNA聚合酶的基因， 细胞和合适的载体，其包含功能性顺反子，其包括外源基因，其侧翼是由RNA聚合酶识别的启动子序列。
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8.发明授权Method for promoting specific alignment of short oligonucleotides on nucleic acids 失效促进短核苷酸对核酸的特异性比对的方法
公开(公告)号：US5547843A
公开(公告)日：1996-08-20
申请号：US325539
申请日：1994-10-18
申请人： F. William Studier , Jan Kieleczawa , John J. Dunn
发明人： F. William Studier , Jan Kieleczawa , John J. Dunn
CPC分类号： C12Q1/6869
摘要： Disclosed is a method for promoting specific alignment of short oligonucleotides on a nucleic acid polymer. The nucleic acid polymer is incubated in a solution containing a single-stranded DNA-binding protein and a plurality of oligonucleotides which are perfectly complementary to distinct but adjacent regions of a predetermined contiguous nucleotide sequence in the nucleic acid polymer. The plurality of oligonucleotides anneal to the nucleic acid polymer to form a contiguous region of double stranded nucleic acid. Specific application of the methods disclosed include priming DNA synthesis and template-directed ligation.
摘要翻译： 公开了一种用于促进短核苷酸在核酸聚合物上的特异性比对的方法。 将核酸聚合物在含有单链DNA结合蛋白和多个与核酸聚合物中预定的连续核苷酸序列的不同但相邻区域完全互补的寡核苷酸的溶液中温育。 多个寡核苷酸与核酸聚合物退火以形成双链核酸的连续区域。 所公开的方法的具体应用包括引发DNA合成和模板定向连接。
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9.发明授权公开(公告)号：US08241887B2
公开(公告)日：2012-08-14
申请号：US12062575
申请日：2008-04-04
申请人： F. William Studier
发明人： F. William Studier
IPC分类号： C12N1/20
摘要： A bacterial growth medium for promoting auto-induction of transcription of cloned DNA in cultures of bacterial cells grown batchwise is disclosed. The transcription is under the control of a lac repressor. Also disclosed is a bacterial growth medium for improving the production of a selenomethionine-containing protein or polypeptide in a bacterial cell, the protein or polypeptide being produced by recombinant DNA techniques from a lac or T7lac promoter, the bacterial cell encoding a vitamin B12-dependent homocysteine methylase. Finally, disclosed is a bacterial growth medium for suppressing auto-induction of expression in cultures of bacterial cells grown batchwise, said transcription being under the control of lac repressor.
摘要翻译： 公开了一种细菌生长培养基，其用于促进分批生长的细菌细胞培养物中克隆DNA的转录自动诱导。 转录受lac阻遏物的控制。 还公开了一种细菌生长培养基，用于改善细菌细胞中含有含硒蛋氨酸的蛋白质或多肽的产生，所述蛋白质或多肽通过来自lac或T7lac启动子的重组DNA技术产生，编码维生素B12依赖性的细菌细胞 高半胱氨酸甲基化酶。 最后，公开了一种细菌生长培养基，其用于抑制细菌生长在分批培养物中的表达的自身诱导，所述转录在lac阻抑物的控制下。
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10.发明授权
公开(公告)号：US5869320A
公开(公告)日：1999-02-09
申请号：US784201
申请日：1997-01-15
申请人： F. William Studier , Parichehre Davanloo , Alan H. Rosenberg , Barbara A. Moffatt , John J. Dunn
发明人： F. William Studier , Parichehre Davanloo , Alan H. Rosenberg , Barbara A. Moffatt , John J. Dunn
CPC分类号： C12N9/1247 , C12N15/70 , C12N15/73
摘要： This application describes a means to clone a functional gene for bacteriophage T7 RNA polymerase. Active T7 RNA polymerase is produced from the cloned gene, and a plasmid has been constructed that can produce the active enzyme in large amounts. T7 RNA polymerase transcribes DNA very efficiently and is highly selective for a relatively long promoter sequence. This enzyme is useful for synthesizing large amounts of RNA in vivo or in vitro, and is capable of producing a single RNA selectively from a complex mixture of DNAs. The procedure used to obtain a clone of the R7 RNA polymerase gene can be applied to other T7-like phages to obtain clones that produce RNA polymerases having different promoter specificities, different bacterial hosts, or other desirable properties. T7 RNA polymerase is also used in a system for selective, high-level synthesis of RNAs and proteins in suitable host cells.
摘要翻译： 本申请描述了克隆噬菌体T7RNA聚合酶的功能基因的方法。 活性T7 RNA聚合酶是从克隆的基因中产生的，已经构建出能大量产生活性酶的质粒。 T7 RNA聚合酶非常有效地转录DNA，对于较长的启动子序列具有高选择性。 该酶可用于在体内或体外合成大量的RNA，并且能够从DNA的复杂混合物中选择性地产生单一RNA。 用于获得R7 RNA聚合酶基因克隆的程序可以应用于其他T7样噬菌体，以获得产生具有不同启动子特异性，不同细菌宿主或其它所需性质的RNA聚合酶的克隆。 T7 RNA聚合酶还用于在合适的宿主细胞中选择性，高水平合成RNA和蛋白质的系统。
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