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1. 发明授权

US5821088A Use of gram-positive bacteria to express recombinant proteins 失效
标题翻译：使用革兰氏阳性细菌表达重组蛋白
公开(公告)号：US5821088A
公开(公告)日：1998-10-13
申请号：US472244
申请日：1995-06-07
申请人： Aldis Darzins , Stephen Whitehead , Dennis Hruby , Vincent A. Fischetti
发明人： Aldis Darzins , Stephen Whitehead , Dennis Hruby , Vincent A. Fischetti
IPC分类号： C12N15/09 , A61K38/00 , A61K39/00 , C07K14/31 , C07K14/315 , C12N15/62 , C12N15/74 , C12P21/02 , C12R1/46 , C12P21/04
CPC分类号： C07K14/31 , C07K14/315 , C12N15/62 , C12N15/74 , A61K38/00 , A61K39/00 , C07K2319/00 , C07K2319/02 , C07K2319/035 , C07K2319/23 , C07K2319/50 , C07K2319/705 , C07K2319/735
摘要： A novel system for cloning and expression of genes in gram-positive bacteria. The expression system is based on the finding that many gram-positive bacteria sort proteins to their cell surface through cis-acting N-terminal signal sequences and C-terminal anchor regions. In particular, the cell sorting signals of the streptococcal M6 protein, a well-known surface molecule, are used to construct a gram-positive expression system, designated SPEX (Streptococcal Protein Expression). Expression is achieved by cloning the gene of interest into an appropriate SPEX cassette which is then stably introduced into a bacterial host, such as the human commensal Streptococcus gordonii. Depending on the SPEX vector used, recombinant proteins can be anchored to the cell wall prior to release by specific endoproteolytic cleavage or secreted into the culture medium during bacterial growth. The use of host bacteria lacking extracellular proteases should protect secreted proteins from proteolytic degradation. Several expression vectors in this system also produce specifically-tagged recombinant proteins which allows for a one-step purification of the resulting product.
摘要翻译：用于克隆和表达革兰氏阳性菌中基因的新系统。表达系统基于许多革兰氏阳性细菌通过顺式作用的N末端信号序列和C末端锚定区域将蛋白质分解到其细胞表面的发现。特别地，使用众所周知的表面分子的链球菌M6蛋白的细胞分选信号来构建称为SPEX（链球菌蛋白表达）的革兰氏阳性表达系统。通过将感兴趣的基因克隆到适当的SPEX盒中，然后将其稳定地导入细菌宿主例如人共生型链球菌（Streptococcus gordonii）中来实现表达。根据所使用的SPEX载体，重组蛋白质可以在通过特异性内切蛋白酶切割释放之前锚定到细胞壁上，或者在细菌生长过程中分泌到培养基中。缺乏细胞外蛋白酶的宿主细菌的使用应保护分泌的蛋白质免于蛋白水解降解。该系统中的几种表达载体也产生特异性标记的重组蛋白，其允许对所得产物进行一步纯化。

2. 发明申请

US20070254934A1 Antiviral drugs for treatment of arenavirus infection 有权
标题翻译：用于治疗斑马鱼病毒感染的抗病毒药物
公开(公告)号：US20070254934A1
公开(公告)日：2007-11-01
申请号：US11712918
申请日：2007-03-02
申请人： Dennis Hruby , Tove Bolken , Sean Amberg , Dongcheng Dai
发明人： Dennis Hruby , Tove Bolken , Sean Amberg , Dongcheng Dai
IPC分类号： A61K31/4164 , A61P31/14 , C07D235/04
CPC分类号： C07D235/06 , A61K31/4164 , Y02A50/387 , Y02A50/397
摘要： Compounds, methods and pharmaceutical compositions for treating viral infections, by administering certain novel compounds in therapeutically effective amounts are disclosed. Methods for preparing the compounds and methods of using the compounds and pharmaceutical compositions thereof are also disclosed. In particular, the treatment and prophylaxis of viral infections such as caused by hemorrhagic fever viruses is disclosed, i.e., including but not limited to, Arenaviridae (Junin, Machupo, Guanarito, Sabia, Lassa, Tacaribe, Pichinde, and LCMV), Filoviridae (Ebola and Marburg viruses), Flaviviridae (yellow fever, Omsk hemorrhagic fever and Kyasanur Forest disease viruses), and Bunyaviridae (Rift Valley fever).
摘要翻译：公开了通过施用治疗有效量的某些新化合物来治疗病毒感染的化合物，方法和药物组合物。还公开了制备化合物的方法和使用该化合物的方法及其药物组合物。具体地，公开了治疗和预防由出血热病毒引起的病毒感染，即，包括但不限于：金杆菌科（Junin，Machupo，Guanarito，Sabia，Lassa，Tacaribe，Pichinde和LCMV），丝状病毒科埃博拉病毒和马尔堡病毒），黄病毒科（黄热病，鄂木斯克出血热和Kyasanur森林病毒）和布亚病毒科（裂谷热）。

3. 发明申请

US20050233396A1 Screening method for orthopoxvirus antivirals 失效
标题翻译：正痘病毒抗病毒药物的筛选方法
公开(公告)号：US20050233396A1
公开(公告)日：2005-10-20
申请号：US10504687
申请日：2003-01-08
申请人： Dennis Hruby , Tove Bolken , Chelsea Byrd
发明人： Dennis Hruby , Tove Bolken , Chelsea Byrd
IPC分类号： G01N33/50 , A61K45/00 , A61P31/12 , A61P43/00 , C12N15/09 , C12Q1/18 , C12Q1/37 , G01N33/15 , G01N33/53 , G01N33/537 , G01N33/542 , G01N33/543 , G01N33/573 , G01N33/68
CPC分类号： G01N33/6842 , C12Q1/18 , C12Q1/37 , G01N33/542 , G01N33/573 , G01N33/6803 , G01N2333/065 , G01N2500/00
摘要： The focus of the present invention is an effective anti-poxvirus drug for use in treating or preventing human disease caused by pathogenic poxviruses. More particularly, the present invention relates to antiviral drugs that target the poxvirus proteinase responsible for core protein maturation, a step which is absolutely essential for the production and spread of infectious virions.
摘要翻译：本发明的重点是用于治疗或预防由病原性痘病毒引起的人类疾病的有效的抗痘病毒药物。更具体地，本发明涉及靶向负责核心蛋白质成熟的痘病毒蛋白酶的抗病毒药物，这是对感染性病毒粒子的产生和扩散绝对必要的步骤。

4. 发明申请

US20050042756A1 Intergenic and intragenic integration sites for foreign gene expression in recombinant S. gordonii strains 审中-公开
标题翻译：外源基因表达在重组的戈登假单胞菌菌株中的基因和基因内整合位点
公开(公告)号：US20050042756A1
公开(公告)日：2005-02-24
申请号：US10698439
申请日：2003-11-03
申请人： Christine Franke , Tove Bolken , Kevin Jones , Dennis Hruby
发明人： Christine Franke , Tove Bolken , Kevin Jones , Dennis Hruby
IPC分类号： C12N1/21 , C12N15/74
CPC分类号： C12N15/74 , C12N15/746
摘要： The present invention provides two new chromosomal integration sites for expression of foreign genes have been developed in Streptococcus gordonii (S. gordonii). One integration site is intergenic between orfA and orfB in an operon of unknown function. The other site is intragenic within the lacG gene, which encodes phospho-β-galactosidase, and is part of the lactose (lac) operon. The emm6 gene from Streptococcus pyogenes was integrated in a stable configuration into the chromosome of S. gordonii at each of these integration sites, and in both cases the recombinant bacteria expressed the M6 protein on their surface. Furthermore, expression from the lacG site within the lactose operon was shown to be regulated by growth on lactose. Identification of these new chromosomal insertion sites provides the ability to express multiple foreign genes from the same recombinant and the potential for modulating expression in vitro or in vivo by the use of a biosynthetic metabolite.
摘要翻译：本发明提供了两种新的染色体整合位点，用于在链球菌（S.gordonii）中表达外源基因。一个整合位点是orfA和orfB在功能未知的操纵子之间的基因。另一个位点是lacG基因内部的基因，其编码磷酸-β-半乳糖苷酶，并且是乳糖（lac）操纵子的一部分。来自化脓性链球菌的emm6基因在这些整合位点的每一个整合到稳定的构型中，并且在这两种情况下，重组细菌在其表面上表达M6蛋白。此外，乳糖操纵子内的lacG位点的表达显示出受乳糖生长的调节。这些新的染色体插入位点的鉴定提供了从相同的重组体表达多个外来基因并通过使用生物合成代谢物在体外或体内调节表达的潜力的能力。

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