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31. 发明授权

US5928908A Method for introducing unidirectional nested deletions 失效
标题翻译：引入单向嵌套缺失的方法
公开(公告)号：US5928908A
公开(公告)日：1999-07-27
申请号：US966958
申请日：1997-11-10
申请人： John J. Dunn , Mark A. Quesada , Matthew Randesi
发明人： John J. Dunn , Mark A. Quesada , Matthew Randesi
IPC分类号： C12N15/10 , C12N15/64
CPC分类号： C12N15/102
摘要： Disclosed is a method for the introduction of unidirectional deletions in a cloned DNA segment. More specifically, the method comprises providing a recombinant DNA construct comprising a DNA segment of interest inserted in a cloning vector, the cloning vector having an f1 endonuclease recognition sequence adjacent to the insertion site of the DNA segment of interest. The recombinant DNA construct is then contacted with the protein pII encoded by gene II of phage f1 thereby generating a single-stranded nick. The nicked DNA is then contacted with E. coli Exonuclease III thereby expanding the single-stranded nick into a single-stranded gap. The single-stranded gapped DNA is then contacted with a single-strand-specific endonuclease thereby producing a linearized DNA molecule containing a double-stranded deletion corresponding in size to the single-stranded gap. The DNA treated in this manner is then incubated with DNA ligase under conditions appropriate for ligation. Also disclosed is a method for producing single-stranded DNA probes. In this embodiment, single-stranded gapped DNA, produced as described above, is contacted with a DNA polymerase in the presence of labeled nucleotides to fill in the gap. This DNA is then linearized by digestion with a restriction enzyme which cuts outside the DNA segment of interest. The product of this digestion is then denatured to produce a labeled single-stranded nucleic acid probe.
摘要翻译：公开了在克隆的DNA片段中引入单向缺失的方法。更具体地，该方法包括提供包含插入克隆载体中的目标DNA片段的重组DNA构建体，该克隆载体具有与感兴趣的DNA片段的插入位点相邻的f1内切核酸酶识别序列。然后将重组DNA构建体与由噬菌体f1的基因II编码的蛋白质pII接触，从而产生单链切口。然后将切口的DNA与大肠杆菌外切核酸酶III接触，从而将单链缺口扩大为单链间隙。然后将单链间隙DNA与单链特异性内切核酸酶接触，从而产生含有与单链间隙大小相对应的双链缺失的线性化DNA分子。然后以适合于连接的条件，用这种方式处理的DNA与DNA连接酶一起温育。还公开了用于生产单链DNA探针的方法。在该实施方案中，如上所述制备的单链间隙DNA在DNA标记的核苷酸的存在下与DNA聚合酶接触以填充间隙。然后通过用限制性内切酶消化DNA直接切割该DNA，该限制性内切酶在感兴趣的DNA片段之外。然后将该消化产物变性以产生标记的单链核酸探针。

32. 发明授权

US5693489A Cloning and expression of the gene for bacteriophage T7 RNA polymerase 失效
标题翻译：噬菌体T7 RNA聚合酶基因的克隆和表达
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
IPC分类号： C12N1/21 , C12N9/12 , C12N15/34 , C12N15/70 , C12N15/73 , C12N15/67 , C12N15/00
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。

33. 发明授权

US5547843A Method for promoting specific alignment of short oligonucleotides on nucleic acids 失效
标题翻译：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
IPC分类号： C12Q1/68 , C12P19/34
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。

34. 发明申请

US20130040343A1 Methods for Detection of Methyl-CpG Dinucleotides 失效
标题翻译：甲基CpG二核苷酸检测方法
公开(公告)号：US20130040343A1
公开(公告)日：2013-02-14
申请号：US13565954
申请日：2012-08-03
申请人： John J. Dunn
发明人： John J. Dunn
IPC分类号： C12P19/34
CPC分类号： C12P19/34 , C07K14/245 , C07K2319/40 , C12P19/36 , C12Q1/6827 , C12Q2523/125 , C12Q2522/101
摘要： The invention provides methods for enriching methyl-CpG sequences from a DNA sample. The method makes use of conversion of cytosine residues to uracil under conditions in which methyl-cytosine residues are preserved. Additional methods of the invention enable to preservation of the context of me-CpG dinucleotides. The invention also provides a recombinant, full length and substantially pure McrA protein (rMcrA) for binding and isolation of DNA fragments containing the sequence 5′-CMeCpGG-3′. Methods for making and using the rMcrA protein, and derivatives thereof are provided.
摘要翻译：本发明提供了从DNA样品中富集甲基CpG序列的方法。该方法利用在保留甲基 - 胞嘧啶残基的条件下将胞嘧啶残基转化成尿嘧啶。本发明的另外的方法能够保存me-CpG二核苷酸的上下文。本发明还提供了用于结合和分离含有5'-CMeCpGG-3'序列的DNA片段的重组全长和基本上纯的McrA蛋白（rMcrA）。提供了制备和使用rMcrA蛋白的方法及其衍生物。

35. 发明授权

US08313691B2 Lean austenitic stainless steel 有权
标题翻译：精益奥氏体不锈钢
公开(公告)号：US08313691B2
公开(公告)日：2012-11-20
申请号：US12037477
申请日：2008-02-26
申请人： David S. Bergstrom , James M. Rakowski , Charles P. Stinner , John J. Dunn , John F. Grubb
发明人： David S. Bergstrom , James M. Rakowski , Charles P. Stinner , John J. Dunn , John F. Grubb
IPC分类号： C22C38/44 , C22C38/58 , C22C38/52
CPC分类号： C22C38/58 , C22C38/001 , C22C38/002 , C22C38/02 , C22C38/22 , C22C38/30 , C22C38/32 , C22C38/34 , C22C38/38 , C22C38/42 , C22C38/44 , C22C38/52 , C22C38/54
摘要： An austenitic stainless steel having low nickel and molybdenum and exhibiting comparable corrosion resistance and formability properties to higher nickel and molybdenum alloys comprises, in weight %, up to 0.20 C, 2.0-9.0 Mn, up to 2.0 Si, 16.0-23.0 Cr, 1.0-5.0 Ni, up to 3.0 Mo, up to 3.0 Cu, 0.1-0.35 N, up to 4.0 W, up to 0.01 B, up to 1.0 Co, iron and impurities, the steel having a ferrite number of less than 10 and a MD30 value of less than 20° C.
摘要翻译：具有低镍和钼的奥氏体不锈钢并且具有与较高镍和钼合金相当的耐腐蚀性和成形性能的奥氏体不锈钢包括以重量％计高达0.20C，2.0-9.0Mn，高达2.0Si，16.0-23.0Cr，1.0 -5.0Ni，高达3.0Mo，至多3.0Cu，0.1-0.35N，至多4.0W，至多0.01B，至多1.0Co，铁和杂质，所述钢的铁素体数小于10， MD30值小于20°C

36. 发明申请

US20090011511A1 Single-Point Genome Signature Tags 审中-公开
标题翻译：单点基因组签名标签
公开(公告)号：US20090011511A1
公开(公告)日：2009-01-08
申请号：US11925382
申请日：2007-10-26
申请人： John J. Dunn , Daniel van der Lelie , Maureen K. Krause , Sean R. McCorkle
发明人： John J. Dunn , Daniel van der Lelie , Maureen K. Krause , Sean R. McCorkle
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6809 , C12Q1/6827 , C12Q2525/191 , C12Q2525/131 , C12Q2521/313 , C12Q2537/159 , C12Q2537/164 , C12Q2563/131
摘要： Disclosed is a method for analyzing the organismic complexity of a sample through analysis of the nucleic acid in the sample. In the disclosed method, through a series of steps, including digestion with a type II restriction enzyme, ligation of capture adapters and linkers and digestion with a type IIS restriction enzyme, genome signature tags are produced. The sequences of a statistically significant number of the signature tags are determined and the sequences are used to identify and quantify the organisms in the sample. Various embodiments of the invention described herein include methods for using single point genome signature tags to analyze the related families present in a sample, methods for analyzing sequences associated with hyper- and hypo-methylated CpG islands, methods for visualizing organismic complexity change in a sampling location over time and methods for generating the genome signature tag profile of a sample of fragmented DNA.
摘要翻译：公开了通过分析样品中的核酸来分析样品的生物复杂性的方法。在所公开的方法中，通过一系列步骤，包括用II型限制酶消化，连接捕获适配器和接头并用IIS限制酶消化，产生基因组签名标签。确定统计学上显着数量的签名标签的序列，并且使用该序列来鉴定和量化样品中的生物体。本文描述的本发明的各种实施方案包括使用单点基因组签名标签分析样品中存在的相关家族的方法，用于分析与超低甲基化和次甲基化CpG岛相关的序列的方法，用于可视化取样中的有机体复杂性变化的方法随着时间推移的位置以及用于产生片段化DNA样品的基因组特征标签谱的方法。

37. 发明授权

US06551420B1 Duplex stainless steel 有权
标题翻译：双相不锈钢
公开(公告)号：US06551420B1
公开(公告)日：2003-04-22
申请号：US09981074
申请日：2001-10-16
申请人： David S. Bergstrom , John J. Dunn , John F. Grubb , William A. Pratt
发明人： David S. Bergstrom , John J. Dunn , John F. Grubb , William A. Pratt
IPC分类号： C22C3844
CPC分类号： C22C38/001 , C21D2211/001 , C21D2211/005 , C22C38/42 , C22C38/44 , C22C38/54 , C22C38/58
摘要： A duplex stainless steel including, in weight percent, up to 0.06 percent carbon, 15 up to less than 25 percent chromium, greater than 3 up to 6 percent nickel, up to 3.75 percent manganese, 0.14 up to 0.35 percent nitrogen, up to 2 percent silicon, greater than 1.4 up to less than 2.5 percent molybdenum, up to less than 0.5 percent copper, up to less than 0.2 percent cobalt, up to 0.05 percent phosphorous, up to 0.005 percent sulfur, and 0.001 up to 0.0035 percent boron, with the remainder being iron and incidental impurities is disclosed. The duplex stainless steel may be included in an article of manufacture, such as a strip, bar, plate, sheet, casting, tubing or piping. A method for making such a duplex stainless steel is also disclosed.
摘要翻译：一种双相不锈钢，包括重量百分比，最多0.06％的碳，15％至少于25％的铬，大于3％至6％的镍，至多3.75％的锰，0.14％至0.35％的氮，至多2 百分比的硅，大于1.4，至少小于2.5％的钼，高达小于0.5％的铜，高达少于0.2％的钴，高达0.05％的磷，至多0.005％的硫，以及0.001至0.0035％的硼，剩下的是铁和附带的杂质。双相不锈钢可以包括在制品中，例如条，条，板，片，铸造，管道或管道。还公开了制造这种双相不锈钢的方法。

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