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121. 发明授权

US06455254B1 Sequence based screening 有权
公开(公告)号：US06455254B1
公开(公告)日：2002-09-24
申请号：US09571499
申请日：2000-05-15
申请人： Jay M. Short
发明人： Jay M. Short
IPC分类号： C12Q168
CPC分类号： C12N15/1034 , C12N15/52 , C12Q1/00 , C12Q1/6811 , C12Q1/6876 , C12Q1/689 , C12Q2600/156 , Y02A90/26
摘要： Provided is a method of obtaining a nucleic acid profile of a sample. The method includes creating a DNA library from a plurality of nucleic acid sequences of a mixed population of organisms and sequencing at least one clone in the DNA library. The sequence is compared to a database and identifying sequences in the database which have homology to a clone in the library thereby obtaining a nucleic acid profile of the mixed population of organisms.

122. 发明授权

US06440668B1 Method of DNA shuffling with polynucleotides produced by blocking or interrupting a synthesis or amplification process 有权
标题翻译：通过阻断或中断合成或扩增过程产生的多核苷酸进行DNA改组的方法
公开(公告)号：US06440668B1
公开(公告)日：2002-08-27
申请号：US09376727
申请日：1999-08-17
申请人： Jay M. Short
发明人： Jay M. Short
IPC分类号： C12Q168
CPC分类号： C12Q1/6811 , C12N15/1027
摘要： Disclosed is a process of performing Sexual PCR which includes generating random polynucleotides by interrupting or blocking synthesis or amplification process to slow or halt synthesis or amplification of at least one polynucleotides, optionally amplifying the polynucleotides, and reannealing the polynucleotides to produce random mutant polynucleotides. Also provided are vector and expression vehicles including such mutant polynucleotides, polypeptides expressed by the mutant polynucleotides and a method for producing random polypeptides.
摘要翻译：公开了进行性PCR的过程，其包括通过中断或阻断合成或扩增过程来产生随机多核苷酸，以减缓或停止至少一种多核苷酸的合成或扩增，任选地扩增多核苷酸，以及重新退火多核苷酸以产生随机突变多核苷酸。还提供了载体和表达载体，包括这种突变体多核苷酸，由突变多核苷酸表达的多肽和用于产生随机多肽的方法。

123. 发明授权

US06368798B1 Screening for novel bioactivities 有权
标题翻译：筛选新生物活性
公开(公告)号：US06368798B1
公开(公告)日：2002-04-09
申请号：US09421970
申请日：1999-10-20
申请人： Jay M. Short
发明人： Jay M. Short
IPC分类号： C12Q168
CPC分类号： C12N9/00 , C12N9/1029 , C12N9/16 , C12N15/102 , C12N15/1034
摘要： Disclosed is a process for identifying clones having a specified enzyme activity by screening for the specified enzyme activity in a library of clones prepared by (i) selectively isolating target nucleic acid from nucleic acid derived from at least one microorganism, by use of at least one polynucleotide probe comprising at least a portion of a nucleic acid sequence encoding an enzyme having the specified enzyme activity; and (ii) transforming a host with isolated target nucleic acid to produce a library of clones which are screened for the specified enzyme activity.
摘要翻译：公开了通过筛选在通过（i）通过使用至少一种微生物从至少一种微生物衍生的核酸中选择性分离靶核酸而制备的克隆文库中筛选特定酶活性来鉴定具有特定酶活性的克隆的方法多核苷酸探针，其包含编码具有特定酶活性的酶的核酸序列的至少一部分; 和（ii）用分离的靶核酸转化宿主以产生筛选特定酶活性的克隆文库。

124. 发明授权

US06358709B1 End selection in directed evolution 有权
标题翻译：定向演化中的终极选择
公开(公告)号：US06358709B1
公开(公告)日：2002-03-19
申请号：US09522289
申请日：2000-03-09
申请人： Jay M. Short , Gerhard Johann Frey
发明人： Jay M. Short , Gerhard Johann Frey
IPC分类号： C12P206
CPC分类号： C07K14/445 , A61K39/00 , A61K2039/53 , C12N9/00 , C12N9/14 , C12N9/16 , C12N9/88 , C12N15/102 , C12N15/1027 , C12N15/1034 , C12Q1/6811 , C12Y301/11002
摘要： This invention provides methods of obtaining novel polynucleotides and encoded polypeptides by the use of non-stochastic methods of directed evolution (DirectEvolution™). A particular advantage of end-selection-based methods is the ability to recover full-length polynucleotides from a library of progeny molecules generated by mutagenesis methods. These methods include non-stochastic polynucleotide site-saturation mutagenesis (Gene Site Saturation Mutagenesis™) and non-stochastic polynucleotide reassembly (GeneReassembly™). This invention provides methods of obtaining novel enzymes that have optimized physical &/or biological properties. Through use of the claimed methods, genetic vaccines, enzymes, small molecules, and other desirable molecules can be evolved towards desirable properties. For example, vaccine vectors can be obtained that exhibit increased efficacy for use as genetic vaccines. Vectors obtained by using the methods can have, for example, enhanced antigen expression, increased uptake into a cell, increased stability in a cell, ability to tailor an immune response, and the like. Furthermore, this invention provides methods of obtaining a variety of novel biologically active molecules, in the fields of antibiotics, pharmacotherapeutics, and transgenic traits.
摘要翻译：本发明提供了通过使用定向进化（DirectEvolution TM）的非随机方法获得新的多核苷酸和编码的多肽的方法。基于终端选择的方法的特别优点是从通过诱变方法产生的后代分子的文库中恢复全长多核苷酸的能力。这些方法包括非随机多核苷酸位点饱和诱变（Gene Site Saturation Mutagenesis TM）和非随机多核苷酸重组（GeneReassembly TM）。本发明提供获得具有优化的物理和/或生物学特性的新型酶的方法。通过使用所要求保护的方法，可以将遗传疫苗，酶，小分子和其它所需分子演变成期望的性质。例如，可以获得表现出增加用作基因疫苗的功效的疫苗载体。通过使用该方法获得的载体可具有例如增强的抗原表达，增加的细胞摄取，增加细胞的稳定性，定制免疫应答的能力等。此外，本发明提供了在抗生素，药物治疗剂和转基因性状领域中获得各种新型生物活性分子的方法。

125. 发明授权

US06335179B1 Directed evolution of thermophilic enzymes 有权
标题翻译：嗜热酶的定向进化
公开(公告)号：US06335179B1
公开(公告)日：2002-01-01
申请号：US09185373
申请日：1998-11-03
申请人： Jay M. Short
发明人： Jay M. Short
IPC分类号： C12P2106
CPC分类号： C12N15/1058 , C12N9/00 , C12N9/16 , C12N9/2405 , C12N9/2468 , C12N9/2471 , C12N15/102 , C12N15/1034 , C12Y302/01021 , C12Y302/01023
摘要： Thermostable enzymes are subjected to mutagenesis to produce a thermophilic enzyme which is stable at thermophilic temperature and which has increased activities at least two-fold higher than the activity of the wild-type thermostable enzyme at lower temperatures, which are temperatures of 50° C. or lower.
摘要翻译：热稳定酶进行诱变以产生在嗜温温度下稳定的嗜热酶，其活性比野生型热稳定酶在50℃温度较低的温度下活性提高至少两倍。或更低。

126. 发明授权

US06280926B1 Gene expression library produced from DNA from uncultivated microorganisms and methods for making the same 失效
标题翻译：从未培养的微生物的DNA产生的基因表达文库及其制备方法
公开(公告)号：US06280926B1
公开(公告)日：2001-08-28
申请号：US08988224
申请日：1997-12-10
申请人： Jay M. Short
发明人： Jay M. Short
IPC分类号： C12Q100
CPC分类号： C12N15/1086 , C12N15/1034 , C12N15/1079 , C12N15/1093 , C12N15/52 , C12Q1/00
摘要： Disclosed is a process of screening clones having DNA from an uncultivated microorganism for a specified protein, e.g. enzyme, activity by screening for a specified protein, e.g. enzyme, activity in a library of clones prepared by (i) recovering DNA from a DNA population derived from at least one uncultivated microorganism; and (ii) transforming a host with recovered DNA to produce a library of clones which is screened for the specified protein, e.g. enzyme, activity.
摘要翻译：公开了一种筛选具有来自未培养的微生物的DNA的克隆用于特定蛋白质的方法，例如，酶，通过筛选特定蛋白质的活性，例如酶，通过（i）从源自至少一种未培养微生物的DNA群体回收DNA的克隆文库中的活性; 和（ii）用回收的DNA转化宿主以产生筛选特定蛋白质的克隆文库，例如。酶，活性。

127. 发明授权

US06232103B1 Methods useful for nucleic acid sequencing using modified nucleotides comprising phenylboronic acid 有权
标题翻译：用于使用包含苯基硼酸的修饰核苷酸进行核酸测序的方法
公开(公告)号：US06232103B1
公开(公告)日：2001-05-15
申请号：US09274711
申请日：1999-03-23
申请人： Jay M. Short
发明人： Jay M. Short
IPC分类号： C12P1934
CPC分类号： C12Q1/6869 , C12Q2527/125
摘要： The present invention relates to a method for sequencing nucleic acids. In particular, such method includes incorporating certain modified nucleotides referred to as Simtides into nucleic acid strands. These Simtides are complexes comprising a nucleotide base (or analog thereof), a linker, and a label capable of generating a detectable signal for sequencing. A Simtide may be incorporated into a sequencing fragment as a primer component, as a chain elongator, or as a chain terminator.
摘要翻译：本发明涉及核酸测序方法。特别地，这种方法包括将称为Simtides的某些修饰的核苷酸掺入核酸链中。这些Simtides是包含核苷酸碱基（或其类似物），接头和能够产生用于测序的可检测信号的标签的复合物。 Simtide可以作为引物组分，作为链增长剂或作为链终止剂掺入到测序片段中。

128. 发明授权

US06168919A Screening methods for enzymes and enzyme kits 有权
标题翻译：酶和酶试剂盒的筛选方法
公开(公告)号：US06168919A
公开(公告)日：2001-01-02
申请号：US08983367
申请日：1998-09-30
申请人： Jay M. Short
发明人： Jay M. Short
IPC分类号： C12Q168
CPC分类号： C40B40/02 , C12N15/1037 , C12N15/1086 , C12N15/1093 , C12N15/52 , C12Q1/00
摘要： Recombinant enzyme libraries and kits where a plurality of enzymes are each characterized by different physical and/or chemical characteristics and classified by common characteristics. The characteristics are determined by screening of recombinant enzymes expressed by a DNA library produced from various microorganisms. Also disclosed is a process for identifying clones of a recombinant library which express a protein with a desired ctivity by screening a library of expression clones randomly produced from DNA of at least one microorganism, said screeing being effected on expression products of said clones to thereby identify clones which express a protein with a desired activity. Also disclosed is a process of screening clones having DNA from an uncultivated microorganism for a specified protein activity by screening for a specified protein activity in a library of clones prepared by (I) recovering DNA from a DNA population derived from at least one uncultivated microorganism; and (ii) transforming a host with recovered DNA to produce a library of clones which is screened for the specified protein activity.
摘要翻译：重组酶文库和试剂盒，其中多个酶各自以不同的物理和/或化学特征为特征，并通过共同特征分类。通过筛选由各种微生物产生的DNA文库表达的重组酶来确定特征。还公开了通过筛选由至少一种微生物的DNA随机产生的表达克隆文库来鉴定表达具有所需作用性的蛋白质的重组文库的克隆的方法，所述克隆在所述克隆的表达产物上进行鉴定，从而鉴定表达具有所需活性的蛋白质的克隆。还公开了通过筛选通过（I）从源自至少一个未培养微生物的DNA群体中回收DNA的克隆文库中筛选特定蛋白质活性来筛选具有来自未培养的微生物的具有DNA的克隆的克隆的方法，以获得特定的蛋白质活性; 和（ii）用回收的DNA转化宿主以产生筛选特定蛋白质活性的克隆文库。

129. 发明授权

US5965408A Method of DNA reassembly by interrupting synthesis 失效
标题翻译：通过中断合成DNA重组的方法
公开(公告)号：US5965408A
公开(公告)日：1999-10-12
申请号：US677112
申请日：1996-07-09
申请人： Jay M. Short
发明人： Jay M. Short
IPC分类号： C12N15/09 , C07K2/00 , C12N15/10 , C12P21/02 , C12Q1/68 , C12P19/34 , C07H21/04 , C12N9/12 , G01N33/48
CPC分类号： C12Q1/6811 , C12N15/1027
摘要： Disclosed is a process of performing Sexual PCR which includes generating random polynucleotides by interrupting or blocking a synthesis or amplification process to show or halt synthesis or amplification of at least one polynucleotide, optionally amplifying the polynucleotides, and reannealing the polynucleotides to produce random mutant polynucleotides. Also provided are vector and expression vehicles including such mutant polynucleotides, polypeptides expressed by the mutant polynucleotides and a method for producing random mutant polypeptides.
摘要翻译：公开了进行性PCR的过程，其包括通过中断或阻断合成或扩增过程来产生随机多核苷酸，以显示或停止至少一种多核苷酸的合成或扩增，任选地扩增多核苷酸以及重新退火多核苷酸以产生随机突变多核苷酸。还提供了载体和表达载体，包括这种突变多核苷酸，由突变多核苷酸表达的多肽和用于产生随机突变多肽的方法。

130. 发明授权

US5958672A Protein activity screening of clones having DNA from uncultivated microorganisms 失效
公开(公告)号：US5958672A
公开(公告)日：1999-09-28
申请号：US657409
申请日：1996-06-03
申请人： Jay M. Short
发明人： Jay M. Short
IPC分类号： C12N15/09 , C12N1/21 , C12N9/00 , C12N15/10 , C12N15/52 , C12Q1/00 , C12Q1/25 , C12Q1/68
CPC分类号： C12N15/1086 , C12N15/1034 , C12N15/1079 , C12N15/1093 , C12N15/52 , C12Q1/00
摘要： Disclosed is a process of screening clones having DNA from an uncultivated microorganism for a specified protein, e.g. enzyme, activity by screening for a specified protein, e.g. enzyme, activity in a library of clones prepared by (i) recovering DNA from a DNA population derived from at least one uncultivated microorganism; and (ii) transforming a host with recovered DNA to produce a library of clones which is screened for the specified protein, e.g. enzyme, activity.

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