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1.发明授权Modification of protein by use of a controllable interveining protein sequence 失效通过使用可控的中间蛋白质序列修饰蛋白质
公开(公告)号：US5496714A
公开(公告)日：1996-03-05
申请号：US4139
申请日：1992-12-09
IPC分类号： C07K14/47 , C12N9/12 , C12N9/22 , C12N9/24 , C12N9/38 , C12N15/62 , C12N15/66 , C12N15/67 , C12P21/00
CPC分类号： C12Y302/01081 , C07K14/47 , C07K14/4716 , C12N15/62 , C12N15/66 , C12N15/67 , C12N9/1252 , C12N9/22 , C12N9/2468 , C12N9/2471 , C12Y301/27009 , C12Y302/01023 , C07K2319/00 , C07K2319/20 , C07K2319/21 , C07K2319/92
摘要： The present invention is directed to modified proteins and methods of their production. The modified proteins comprise a controllable intervening protein sequence (CIVPS) inserted into a target protein, the CIVPS being capable of excision from the modified protein under predetermined conditions, i.e., increase in temperature, exposure to light, unblocking of amino acid residues by dephosphorylation or deglycosylation. If desired, the modified protein can be subjected to these conditions. The CIVPS may also be inserted into a region that substantially inactivates target protein activity.
摘要翻译： 本发明涉及修饰的蛋白质及其生产方法。 修饰的蛋白质包含插入目标蛋白质的可控的插入蛋白质序列（CIVPS），CIVPS能够在预定条件下从修饰的蛋白质中切除，即温度升高，暴露于光，通过去磷酸化解除氨基酸残基的阻断或 去糖基化。 如果需要，修饰的蛋白质可以经受这些条件。 CIVPS也可以插入基本上灭活靶蛋白活性的区域。
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2.发明授权Modified proteins comprising controllable intervening protein sequences or their elements methods of producing same and methods for purification of a target protein comprised by a modified protein 失效包含可控中间蛋白序列或其元件的修饰蛋白质的制备方法以及由修饰的蛋白质组成的靶蛋白的纯化方法
公开(公告)号：US5834247A
公开(公告)日：1998-11-10
申请号：US811492
申请日：1997-03-05
申请人： Donald G. Comb , Francine B. Perler , William E. Jack , Ming-Qun Xu , Robert A. Hodges , Christopher J. Noren , Shaorong S. C. Chong , Eric Adam , Maurice Southworth
发明人： Donald G. Comb , Francine B. Perler , William E. Jack , Ming-Qun Xu , Robert A. Hodges , Christopher J. Noren , Shaorong S. C. Chong , Eric Adam , Maurice Southworth
IPC分类号： C07K1/107 , C07K14/47 , C12N9/12 , C12N9/14 , C12N9/22 , C12N9/24 , C12N9/38 , C12N15/62 , C12N15/66 , C12N15/67 , C12P21/02 , C07K19/00 , C12P21/00 , C12P21/04
CPC分类号： C12Y302/01081 , C07K1/1075 , C07K14/47 , C07K14/4716 , C12N15/62 , C12N15/66 , C12N15/67 , C12N9/1252 , C12N9/14 , C12N9/22 , C12N9/2471 , C12P21/02 , C12Y302/01023 , C07K2319/00 , C07K2319/20 , C07K2319/21 , C07K2319/92 , C07K2319/95
摘要： The present invention is directed to modified proteins and methods of their production. The modified proteins comprise a controllable intervening protein sequence (CIVPS) inserted into or adjacent a target protein, the CIVPS being capable of excision from or cleavage of the modified protein under predetermined conditions in cis or in trans, i.e., increase in temperature, exposure to light, unblocking of amino acid residues by dephosphorylation, treatment with chemical reagents or deglycosylation. If desired, the modified protein can be subjected to these conditions. The CIVPS may also be inserted into a region that substantially inactivates target protein activity. The CIVPS may be used in a number of applications including purification of the target protein in a one-step protocol.
摘要翻译： 本发明涉及修饰的蛋白质及其生产方法。 修饰的蛋白质包含插入或邻近靶蛋白质的可控插入蛋白质序列（CIVPS），CIVPS能够在预定条件下以顺式或反式切割或切割修饰的蛋白质，即温度升高，暴露于 通过去磷酸化解除氨基酸残基，用化学试剂或去糖基化处理。 如果需要，修饰的蛋白质可以经受这些条件。 CIVPS也可以插入基本上灭活靶蛋白活性的区域。 CIVPS可用于许多应用，包括以一步方案纯化靶蛋白。
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公开(公告)号：US5834285A
公开(公告)日：1998-11-10
申请号：US222715
申请日：1994-04-04
CPC分类号： C07K16/12 , C12N9/1252
摘要： Recombinant DNA polymerases from archaebacteria as well as isolated DNA coding for such polymerases are provided. The isolated DNA is obtained by use or DNA or antibody probes prepared from the DNA encoding T. litoralis DNA polymerase and the T. litoralis DNA polymerase respectively. Also provided are methods for producing recombinant archaebacteria thermostable DNA polymerase and methods for enhancing the expression of such polymerases by identifying, locating and removing introns from within the DNA coding for such DNA polymerases.
摘要翻译： 提供了来自古细菌的重组DNA聚合酶以及编码这种聚合酶的分离的DNA。 分离的DNA是通过使用DNA或抗体探针获得的，分别是DNA编码的DNA或抗体，分别由编码泰氏针茅DNA和聚合酶链反应的DNA聚合酶。 还提供了用于产生重组古细菌耐热性DNA聚合酶的方法和通过鉴定，定位和从编码这种DNA聚合酶的DNA内鉴定和去除内含子来增强这些聚合酶的表达的方法。
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公开(公告)号：US5352778A
公开(公告)日：1994-10-04
申请号：US167238
申请日：1993-12-15
CPC分类号： C07K16/12 , C12N9/1252
摘要： Recombinant DNA polymeraset from archaebacteria as well as isolated DNA coding for such polymeraset are provided. The isolated DNA is obtained by use of DNA or antibody probet prepared from the DNA encoding T. litoralis DNA polymerate and the T. litoralis DNA polymerate respectively. Also provided are meshocs for producing recombinant archaebacteria thermostable DNA polymerase and methods for enhancing the expression of such polymeraset by identifying, locating and removing introns from within the DNA coding for such DNA polymerases.
摘要翻译： 提供了来自古细菌的重组DNA聚合酶以及编码这种聚合酶的分离的DNA。 分离的DNA通过使用分别由编码泰氏单胞菌DNA聚合物的DNA和泰氏单胞菌DNA聚合物制备的抗体探针获得。 还提供了用于产生重组古细菌耐热性DNA聚合酶的方法和通过从编码这种DNA聚合酶的DNA内鉴定，定位和去除内含子来增强这些聚合酶的表达的方法。
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5.发明授权
公开(公告)号：US5322785A
公开(公告)日：1994-06-21
申请号：US686340
申请日：1991-04-17
IPC分类号： C12N9/10 , C12N1/21 , C12N9/12 , C12N9/16 , C12N15/09 , C12N15/54 , C12N15/55 , C12R1/01 , C12R1/19 , C12N15/67 , C12N15/70 , C12N15/74
CPC分类号： C12N9/1252
摘要： There is provided an extremely thermostable enzyme obtainable from Thermococcus litoralis. The thermostable enzyme has a molecular weight of about 90,000-95,000 daltons, a half-life of about 60 minutes at 100.degree. C. in the absence of stabilizer, and a half-life of about 95 minutes at 100.degree. C. in the presence of stabilizer, such as octoxynol (TRITON X-100) or bovine serum albumin. The thermostable enzyme possesses a 3'-5' proofreading exonuclease activity. The thermostable enzyme may be native or recombinant and may be used for second-strand cDNA synthesis in cDNA cloning, DNA sequencing, and DNA amplification.
摘要翻译： 提供了可从Thermococcus litoralis获得的极热稳定的酶。 耐热酶的分子量约为90,000-95,000道尔顿，在不存在稳定剂的情况下，在100℃下半衰期约为60分钟，而在100℃存在时，半衰期约为95分钟 的稳定剂如辛苯醇（TRITON X-100）或牛血清白蛋白。 耐热酶具有3'-5'校对核酸外切酶活性。 耐热酶可以是天然的或重组的，并且可以用于cDNA克隆，DNA测序和DNA扩增中的第二链cDNA合成。
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公开(公告)号：US5500363A
公开(公告)日：1996-03-19
申请号：US117491
申请日：1993-09-07
CPC分类号： C07K16/12 , C12N9/1252
摘要： Recombinant DNA polymerases from archaebacteria as well as isolated DNA coding for such polymerases are provided. The isolated DNA is obtained by use of DNA or antibody probes prepared from the DNA encoding T. litoralis DNA polymerase and the T. litoralis DNA polymerase respectively. Also provided are methods for producing recombinant archaebacteria thermostable DNA polymerase and methods for enhancing the expression of such polymerases by identifying, locating and removing introns from within the DNA coding for such DNA polymerases.
摘要翻译： 提供了来自古细菌的重组DNA聚合酶以及编码这种聚合酶的分离的DNA。 分离的DNA是通过分别使用由编码泰氏单胞菌DNA聚合酶和泰氏单胞菌DNA聚合酶的DNA制备的DNA或抗体探针获得的。 还提供了用于产生重组古细菌耐热性DNA聚合酶的方法和通过鉴定，定位和从编码这种DNA聚合酶的DNA内鉴定和去除内含子来增强这些聚合酶的表达的方法。
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公开(公告)号：US20110045489A1
公开(公告)日：2011-02-24
申请号：US12988729
申请日：2009-04-20
申请人： Andrew Gardner , Lucia Greenough , William E. Jack
发明人： Andrew Gardner , Lucia Greenough , William E. Jack
CPC分类号： C12Y207/07007 , C12N9/1252 , C12P19/34
摘要： Compositions and methods are provided that relate to a recombinant protein with DNA polymerase activity in which one or more amino acids are mutated compared with the corresponding wild type protein. The recombinant protein is capable of incorporating one or more modified nucleotides into a nucleic acid substrate with a specific activity greater than 200.
摘要翻译： 提供了与具有DNA聚合酶活性的重组蛋白质相关的组合物和方法，其中一个或多个氨基酸与相应的野生型蛋白质相比是突变的。 重组蛋白能够将一个或多个修饰的核苷酸掺入特异活性大于200的核酸底物中。
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公开(公告)号：US06660475B2
公开(公告)日：2003-12-09
申请号：US09738444
申请日：2000-12-15
IPC分类号： C12Q168
CPC分类号： C12N15/10 , C12N15/101 , C12N15/102 , C12N15/64 , Y10T436/143333
摘要： The present invention relates to the use of site-specific nucleic acid nicking enzymes to create single-stranded regions in duplex nucleic acids. Such single-stranded regions can take the form of gaps interior to the duplex, or terminal single-stranded regions. Single-stranded termini can be crafted to allow linkage of various elements via base-pairing with elements containing a complementary single-stranded region. This joining is useful, for example, in an ordered, oriented assembly of DNA modules to create cloning or expression vectors. This joining is also useful in attaching detection probes and purifying DNA molecules containing the single-stranded region. Gaps are useful in similar applications, including attaching detection or purification probes.
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