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    System useful for the production of proteins from recombinant DNA in single celled organisms
    1.
    发明授权
    System useful for the production of proteins from recombinant DNA in single celled organisms 失效
    用于从单细胞生物体中重组DNA生产蛋白质的系统

    公开(公告)号：US5942421A

    公开(公告)日：1999-08-24

    申请号：US972294

    申请日：1997-11-18

    申请人： Michael L. Shuler ,  David B. Wilson

    发明人： Michael L. Shuler ,  David B. Wilson

    IPC分类号： C12N15/75 ,  C12P21/00

    CPC分类号： C12N15/75

    摘要： A single celled organism, vector, and method are provided for continuous production of excreted proteins in the absence of substantial host cell replication and without the necessity for the addition of antibiotics to control cell replication. The vector has (1) an inducible promoter capable of activating a gene for a protein to be produced under conditions that substantially inhibit host cell replication, and (2) a hybrid gene containing a signal sequence fused to a gene for the protein to be produced. Large quantities of the transformed single celled organisms containing the vector can be grown in the absence of inducing conditions, thereby reducing the problem of plasmid loss. Further, the protein is produced under conditions that substantially inhibit host cell replication, thereby allowing immobilization of the single celled organism by entrapment or attachment within or onto a solid support surface. The single celled organism can be immobilized after, before or simultaneously with induction. The single celled organism advantageously may be an encapsulated gram-negative bacterium which has no outer membrane or an outer membrane that is permeable to the proteins to be produced so that the protein products are excreted into the media thereby avoiding difficulty in recovering the desired protein products due to the formation of inclusion bodies or the presence of contaminants produced by cell lyses.

    摘要翻译： 提供单细胞生物体，载体和方法用于在不存在显着的宿主细胞复制的情况下连续产生排泄的蛋白质，而不需要加入抗生素来控制细胞复制。 该载体具有（1）能够在基本上抑制宿主细胞复制的条件下激活待产生的蛋白质的基因的诱导型启动子，和（2）含有与要产生的蛋白质的基因融合的信号序列的杂交基因 。 含有载体的大量转化的单细胞生物体可以在不存在诱导条件的情况下生长，从而减少质粒损失的问题。 此外，蛋白质在基本上抑制宿主细胞复制的条件下产生，从而允许单细胞生物体通过捕获或附着固定在固体支持物表面内或附着在固体支持物表面上。 单细胞生物体可以在诱导后，之前或同时被固定。 单细胞有机体有利地可以是包封的革兰氏阴性细菌，其不具有对要产生的蛋白质可渗透的外膜或外膜，使得蛋白质产物被排泄到培养基中，从而避免难以回收所需的蛋白质产物 由于包涵体的形成或细胞溶解产生的污染物的存在。

    System useful for the production of proteins from recombinant DNA in single celled organisms
    2.
    发明授权
    System useful for the production of proteins from recombinant DNA in single celled organisms 失效

    公开(公告)号：US5747281A

    公开(公告)日：1998-05-05

    申请号：US472485

    申请日：1995-06-07

    申请人： Michael L. Shuler ,  David B. Wilson

    发明人： Michael L. Shuler ,  David B. Wilson

    IPC分类号： C12N15/75 ,  C12P21/00 ,  C12N15/00

    CPC分类号： C12N15/75

    摘要： A single celled organism, vector, and method are provided for continuous production of excreted proteins in the absence of substantial host cell replication and without the necessity for the addition of antibiotics to control cell replication. The vector has (1) an inducible promoter capable of activating a gene for a protein to be produced under conditions that substantially inhibit host cell replication, and (2) a hybrid gene containing a signal sequence fused to a gene for the protein to be produced. Large quantities of the transformed single celled organisms containing the vector can be grown in the absence of inducing conditions, thereby reducing the problem of plasmid loss. Further, the protein is produced under conditions that substantially inhibit host cell replication, thereby allowing immobilization of the single celled organism by entrapment or attachment within or onto a solid support surface. The single celled organism can be immobilized after, before or simultaneously with induction. The single celled organism advantageously may be an encapsulated gram-negative bacterium which has no outer membrane or an outer membrane that is permeable to the proteins to be produced so that the protein products are excreted into the media thereby avoiding difficulty in recovering the desired protein products due to the formation of inclusion bodies or the presence of contaminants produced by cell lyses.

    Method and apparatus for managing financial accounts
    3.
    发明授权
    Method and apparatus for managing financial accounts 有权
    管理财务账目的方法和设备

    公开(公告)号：US08239298B1

    公开(公告)日：2012-08-07

    申请号：US10626885

    申请日：2003-07-24

    申请人： David B. Wilson ,  Frederick C. Buffum

    发明人： David B. Wilson ,  Frederick C. Buffum

    IPC分类号： G06Q40/00

    CPC分类号： G06Q40/00

    摘要： Method, system, and computer-readable medium are provided for optimizing an allocation of money to assets and liabilities. A method includes providing at least one asset comprising a value and a projected return rate at which the value is expected to appreciate over a period, providing at least one liability including a liability amount and an interest rate at which the liability amount is expected to depreciate over said period, and optimizing an allocation of money to the at least one asset and the at least one liability based on the expected appreciation of the at least one asset and the expected depreciation of the at least one liability.

    摘要翻译： 提供了方法，系统和计算机可读介质，用于优化资产和负债的分配。 一种方法包括提供至少一种资产，其中包括价值预期在一段时间内升值的价值和预计收益率，提供至少一个负债，包括负债金额和负债金额预期贬值的利率 并且根据至少一项资产的预期升值和至少一项负债的预计折旧来优化对至少一项资产的资金分配和至少一项负债。

    Nucleotide sequences encoding a thermostable alkaline protease
    4.
    发明授权
    Nucleotide sequences encoding a thermostable alkaline protease 失效
    编码热稳定性碱性蛋白酶的核苷酸序列

    公开(公告)号：US5705379A

    公开(公告)日：1998-01-06

    申请号：US736361

    申请日：1996-10-23

    申请人： David B. Wilson ,  Guifang Lao

    发明人： David B. Wilson ,  Guifang Lao

    IPC分类号： C12N9/52 ,  C07H21/04 ,  C12N1/20 ,  C12N9/50

    CPC分类号： C12N9/52

    摘要： Nucleotide sequences, derived from a thermophilic actinomycete microorganism, which encode a thermostable alkaline protease are disclosed. Also disclosed are variants of the nucleotide sequences which encode a polypeptide having thermostable alkaline proteolytic activity. Recombinant thermostable alkaline protease or recombinant polypeptide may be obtained by culturing in a medium a host cell genetically engineered to contain and express a nucleotide sequence according to the present invention, and recovering the recombinant thermostable alkaline protease or recombinant polypeptide from the culture medium.

    摘要翻译： 公开了衍生自嗜热放线菌微生物的编码热稳定性碱性蛋白酶的核苷酸序列。 还公开了编码具有耐热性碱性蛋白水解活性的多肽的核苷酸序列的变体。 重组热稳定性碱性蛋白酶或重组多肽可以通过在基因工程改造成含有和表达本发明的核苷酸序列的宿主细胞的培养基中培养并从培养基中回收重组热稳定性碱性蛋白酶或重组多肽来获得。

    Alkali and heat stable protease from thermomonospora fusca
    8.
    发明授权
    Alkali and heat stable protease from thermomonospora fusca 失效
    碱性热稳定蛋白酶

    公开(公告)号：US5192677A

    公开(公告)日：1993-03-09

    申请号：US713602

    申请日：1991-06-11

    申请人： John E. Kinsella ,  Todd W. Gusek ,  David B. Wilson

    发明人： John E. Kinsella ,  Todd W. Gusek ,  David B. Wilson

    IPC分类号： C12N9/52

    CPC分类号： C12N9/52

    摘要： An exocellular protease from Thermomonospora fusca YX and a process for producing the protease which has the following physicochemical properties:(1) Molecular mass:The protease has a molecular mass of from about 10,000 to 14,000 Daltons as measured by SDS-polyacrylamide gel electrophoresis;(2) Influence of inhibitors:The protease activity is inhibited by serine protease inhibitors;(3) Substrate specificity:A non-specific protease which can hydrolyze food proteins and bovine serum albumin at a rate of 50-100 nmoles peptide bonds/.mu.g enzyme/minute at 55.degree., pH 8.5 in 0.5 M Tris buffer without showing any substrate and/or product inhibition;(4) Reactivity:A broad spectrum serine type protease having activity at least 5 times greater than trypsin or chymotrypsin towards food grade proteins and bovine serum albumin;(5) Optimum activity temperature and temperature range:The optimum activity temperature is 80.degree. C. at pH of 8.0 in 0.05 M Tris buffer at an ionic strength of 0.2 M NaCl; The temperature range is 35.degree. to 95.degree. C. under the same pH, buffer, and ionic strength;(6) pH range and optimum pH value:The protease has a pH activity range of from about 7 to 11, and the optimum pH value is 9.0;(7) Tolerance to ionic strength conditions and optimum ionic strength:The protease is tolerant to ionic strengths of from 0.0 to about 1.0 M NaCl; Optimum ionic strength is 0.2 M NaCl;(8) Isoelectric point:The isoelectric point is at an alkaline pH; and(9) Structure:The protease is a monomer.

    摘要翻译： 一种来自热单孢菌YX的外源细胞蛋白酶和具有以下物理化学性质的蛋白酶的制备方法：（1）分子量：通过SDS-聚丙烯酰胺凝胶电泳测定，蛋白酶的分子量为约10,000-14,000道尔顿; （2）抑制剂的影响：蛋白酶活性被丝氨酸蛋白酶抑制剂抑制; （3）底物特异性：可以在0.5M Tris缓冲液中以50-100nmol肽键/μg酶/分钟的速率在55℃，pH8.5下以50-100nmole肽键/μg酶/分钟的速率水解食物蛋白质和牛血清白蛋白的非特异性蛋白酶， 底物和/或产物抑制; （4）反应性：一种广谱丝氨酸型蛋白酶，其活性比胰蛋白酶或糜蛋白酶大于食品级蛋白质和牛血清白蛋白的5倍以上; （5）最佳活性温度和温度范围：在离子强度为0.2M NaCl的0.05M Tris缓冲液中，最适活性温度为80℃，pH 8.0; 在相同pH，缓冲液和离子强度下，温度范围为35〜95℃; （6）pH范围和最适pH值：蛋白酶的pH值范围为约7〜11，最适pH值为9.0; （7）对离子强度条件和最佳离子强度的耐受性：蛋白酶耐受0.0至约1.0M NaCl的离子强度; 最佳离子强度为0.2M NaCl; （8）等电点：等电点处于碱性pH; 和（9）结构：蛋白酶是单体。

    Clampless anastomotic device
    9.
    发明授权
    Clampless anastomotic device 失效
    无缝吻合装置

    公开(公告)号：US08052639B2

    公开(公告)日：2011-11-08

    申请号：US12099650

    申请日：2008-04-08

    申请人： David B. Wilson

    发明人： David B. Wilson

    IPC分类号： A61M29/00

    CPC分类号： A61B17/11 ,  A61B17/12045 ,  A61B17/12136 ,  A61B2017/1107 ,  A61B2017/1135 ,  A61B2017/12127

    摘要： Described is a medical device and method for allowing fenestration of the aortic wall while maintaining distal perfusion and preventing external bleeding. The device isolates a segment of the aortic wall from the flowing column of blood using a balloon mounted on a metal alloy strut assembly. The strut assembly expands radially from a collapsed, low-profile configuration when uncovered by a constraining outer sheath. Aortic blood flow is allowed through the flow passage thus contained by the strut assembly within the center of the balloon. The balloon is inflated to contact the aortic wall. The balloon contains a pocket shaped to allow aortic fenestration. The balloon contains radiopaque markers to facilitate orientation and positioning of the pocket within the aorta. Other embodiments using spaced balloons are also disclosed.

    摘要翻译： 描述了一种用于允许开启主动脉壁同时保持远端灌注并防止外部出血的医疗装置和方法。 该装置使用安装在金属合金支柱组件上的球囊将主动脉壁的一段与流动的血液柱隔离。 当被约束外护套覆盖时，支柱组件从折叠的低轮廓构造径向膨胀。 允许主动脉血流通过由球囊中心内的支柱组件包含的流动通道。 气囊膨胀以接触主动脉壁。 气囊包含一个口袋形状以允许主动脉开窗。 气球包含不透射线的标记，以便于主动脉内袋的取向和定位。 还公开了使用间隔球囊的其它实施例。

    CLAMPLESS ANASTOMOTIC DEVICE
    10.
    发明申请
    CLAMPLESS ANASTOMOTIC DEVICE 失效
    无夹紧分析装置

    公开(公告)号：US20080255506A1

    公开(公告)日：2008-10-16

    申请号：US12099650

    申请日：2008-04-08

    申请人： David B. Wilson

    发明人： David B. Wilson

    IPC分类号： A61M25/10 ,  A61M25/095 ,  A61M29/00

    CPC分类号： A61B17/11 ,  A61B17/12045 ,  A61B17/12136 ,  A61B2017/1107 ,  A61B2017/1135 ,  A61B2017/12127

    摘要： Described is a medical device and method for allowing fenestration of the aortic wall while maintaining distal perfusion and preventing external bleeding. The device isolates a segment of the aortic wall from the flowing column of blood using a balloon mounted on a metal alloy strut assembly. The strut assembly expands radially from a collapsed, low-profile configuration when uncovered by a constraining outer sheath. Aortic blood flow is allowed through the flow passage thus contained by the strut assembly within the center of the balloon. The balloon is inflated to contact the aortic wall. The balloon contains a pocket shaped to allow aortic fenestration. The balloon contains radiopaque markers to facilitate orientation and positioning of the pocket within the aorta. Other embodiments using spaced balloons are also disclosed.

    摘要翻译： 描述了一种用于允许开启主动脉壁同时保持远端灌注并防止外部出血的医疗装置和方法。 该装置使用安装在金属合金支柱组件上的球囊将主动脉壁的一段与流动的血液柱隔离。 当被约束外护套覆盖时，支柱组件从折叠的低轮廓构造径向膨胀。 允许主动脉血流通过由球囊中心内的支柱组件包含的流动通道。 气囊膨胀以接触主动脉壁。 气囊包含一个口袋形状以允许主动脉开窗。 气球包含不透射线的标记，以便于主动脉内袋的取向和定位。 还公开了使用间隔球囊的其它实施例。