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1. 发明申请

US20120202249A1 PRODUCTION OF AN INTACT VIRUS IN A MAMMALIAN (NON-HOST) CELL SYSTEM USING A SECONDARY NON-HOST VIRAL CONSTRUCT 审中-公开
标题翻译：使用二次非死亡病毒结构在马萨诸塞（非主机）细胞系统中产生致病病毒
公开(公告)号：US20120202249A1
公开(公告)日：2012-08-09
申请号：US13263360
申请日：2010-04-08
申请人： Arun K. Dhar
发明人： Arun K. Dhar
IPC分类号： C12P21/00 , C12N7/00 , C12N5/10 , C12N15/86 , C12N15/866
CPC分类号： C12N15/86 , C12N7/00 , C12N2710/14144 , C12N2770/16043 , C12N2770/24243 , C12N2770/32043 , C12N2800/50 , C12N2830/60
摘要： The present invention relates to constructs and methods for the production of recombinant proteins, viruses, and viral vaccines in heterologous culture systems by expressing intact genes or viral genomes under the control of a pantropic promoter in a culture system that is not considered a host to the virus so produced. The promoter/viral genome constructs are inserted into a baculovirus and expressed in mammalian non-host cells for the baculovirus.
摘要翻译：本发明涉及用于在异源培养系统中生产重组蛋白质，病毒和病毒疫苗的构建体和方法，其通过在不被认为是宿主的培养系统中的泛素启动子控制下表达完整的基因或病毒基因组病毒如此生产。将启动子/病毒基因组构建体插入杆状病毒中，并在用于杆状病毒的哺乳动物非宿主细胞中表达。

2. 发明申请

US20100092521A1 ANTIGENICITY OF INFECTIOUS PANCREATIC NECROSIS VIRUS VP2 SUB-VIRAL PARTICLES EXPRESSED IN YEAST 有权
标题翻译：感染性胰腺炎病毒VP2亚历山大小肠病毒的抗生素
公开(公告)号：US20100092521A1
公开(公告)日：2010-04-15
申请号：US12519991
申请日：2007-12-18
申请人： Arun K. Dhar , Robert M. Bowers , F.C. Thomas Allnutt
发明人： Arun K. Dhar , Robert M. Bowers , F.C. Thomas Allnutt
IPC分类号： A61K39/00 , C12N1/19 , C12P21/00 , A61P31/12
CPC分类号： A61K39/12 , A61K2039/51 , A61K2039/5258 , A61K2039/542 , A61K2039/552 , C07K14/005 , C12N7/00 , C12N2720/10022 , C12N2720/10023 , C12N2720/10034
摘要： Infectious pancreatic necrosis virus (IPNV), the etiologic agent of infectious pancreatic necrosis in salmonid fish, causes significant losses to the aquaculture industry. The gene for the viral capsid protein (VP2) was cloned into a yeast expression vector and expressed in Saccharomyces cerevisae. Expression of the capsid gene in yeast resulted in formation of approximately 20 nanometer sub-viral particles composed solely of VP2 protein. Anti-IPNV antibodies were detected in rainbow trout vaccinated either by injection of purified VP2-subviral particles (rVP2-SVP) or by feeding recombinant yeast expressing rVP2-SVP. Challenge of rVP2-SVP immunized trout with a heterologous IPNV strain and subsequent viral load determination showed that both injection and orally vaccinated fish had lower IPNV loads than naive or sham-vaccinated fish. This study demonstrates the ability of rVP2-SVPs to induce a specific immune response and the ability of immunized fish to reduce the viral load after an experimentally induced IPNV infection. The invention is not limited to IPNV, and is applicable to other similar viruses for which SVPs can be made and administered to fish.
摘要翻译：传染性胰腺坏死病毒（IPNV）是鲑鱼感染性胰腺坏死的病原体，对水产养殖业造成重大损失。将病毒衣壳蛋白（VP2）的基因克隆到酵母表达载体中并在酿酒酵母中表达。酵母中衣壳基因的表达导致形成仅由VP2蛋白组成的约20纳米亚病毒颗粒。通过注射纯化的VP2-亚病毒颗粒（rVP2-SVP）或通过加入表达rVP2-SVP的重组酵母，在虹鳟中检测到抗IPNV抗体。用异源IPNV株进行rVP2-SVP免疫鳟鱼的挑战，随后进行病毒载量测定，结果表明注射和口服接种的鱼类比初始或假手术的鱼具有较低的IPNV负荷。该研究证明rVP2-SVP诱导特异性免疫应答的能力以及免疫鱼在实验诱导的IPNV感染后降低病毒载量的能力。本发明不限于IPNV，适用于可以制造和施用SVP的其他类似病毒。

3. 发明申请

US20140314858A1 IPNV-ISAV BIVALENT VACCINE USING A VIRUS-LIKE PARTICLE-BASED PLATFORM AND METHODS OF USING THE SAME 有权
标题翻译：使用基于病毒颗粒的平台的IPNV-ISAV双重疫苗及其使用方法
公开(公告)号：US20140314858A1
公开(公告)日：2014-10-23
申请号：US14343666
申请日：2012-09-07
申请人： Arun K. Dhar
发明人： Arun K. Dhar
IPC分类号： A61K39/295 , C07K5/103 , C12N7/00 , C07K7/06
CPC分类号： A61K39/295 , A61K39/12 , A61K2039/523 , A61K2039/542 , A61K2039/552 , A61K2039/55566 , A61K2039/70 , C07K5/101 , C07K7/06 , C07K2319/00 , C12N7/00 , C12N2720/10023 , C12N2720/10034 , C12N2720/10044 , C12N2720/12023 , C12N2720/12034 , C12N2760/16034 , C12N2760/16043
摘要： The present invention provides bivalent oral vaccines against infectious pancreatic necrosis virus (IPNV) and infectious salmon anemia virus (ISAV). Yeast cells comprise an expression vector comprising (i) a polynucleotide sequence encoding a VP2 capsid protein of IPNV and (ii) a polynucleotide sequence encoding one or more antigenic epitopes of hemaglutinin of ISAV. The yeast cells express sub-viral particles (SVP) comprising a VP2 capsid protein of IPNV and one or more antigenic epitopes of hemaglutinin of ISAV. The yeast cells and SVP may be administered in an effective amount to increase the amount of antibodies against IPNV and ISAV in the fish, preferably Salmonidae. The yeast cells and SVP may be in the form of fish food for administering to the fish orally.
摘要翻译：本发明提供针对感染性胰腺坏死病毒（IPNV）和感染性鲑鱼贫血病毒（ISAV）的二价口服疫苗。酵母细胞包含表达载体，其包含（i）编码IPNV的VP2衣壳蛋白的多核苷酸序列和（ii）编码ISAV血凝素的一个或多个抗原表位的多核苷酸序列。酵母细胞表达包含IPNV的VP2衣壳蛋白和ISAV的血凝素的一个或多个抗原表位的亚病毒颗粒（SVP）。可以以有效量施用酵母细胞和SVP以增加鱼，优选鲑科的IPNV和ISAV抗体的量。酵母细胞和SVP可以是鱼类的形式，用于口服给鱼。

4. 发明申请

US20110269225A1 CRUSTACEAN EXPRESSION VECTOR 审中-公开
标题翻译：表达式表达向量
公开(公告)号：US20110269225A1
公开(公告)日：2011-11-03
申请号：US13176026
申请日：2011-07-05
申请人： ARUN K. DHAR , F.C. THOMAS ALLNUTT
发明人： ARUN K. DHAR , F.C. THOMAS ALLNUTT
IPC分类号： C12N15/85
CPC分类号： C12N15/85 , A01K2227/70 , A01K2267/0393 , C12N2799/021 , C12N2800/90
摘要： Methods and constructs for genetic manipulation of one or more of shrimp, shellfish, mollusks, and fish are disclosed. The nucleic acid construct includes a promoter and an internal ribosome entry site of an insect picomavirus, such as a cricket paralysis-like picomavirus. One or more open reading frames can be operably associated with one or both of the promoter and the internal ribosome entry site, and one or more proteins or protein subunits can be expressed upon introduction of the construct into a host cell, such as into a shrimp. Method for producing immortalized crustacean cell lines using enhancer elements derived from shrimp and/or shrimp viruses are also described.
摘要翻译：公开了一种或多种虾，贝类，软体动物和鱼的遗传操作的方法和构造。核酸构建体包括一种启动子和一种诸如板球麻痹状皮莫瓦病毒之类的皮肤微病毒病毒的内部核糖体进入位点。一个或多个开放阅读框可以与启动子和内部核糖体进入位点中的一个或两个可操作地相关联，并且一个或多个蛋白质或蛋白质亚基可以在将构建体引入宿主细胞时表达，例如进入虾。还描述了使用源自虾和/或虾病毒的增强子元件生产永生化甲壳类细胞系的方法。

5. 发明申请

US20090181363A1 NON-INVASIVE DETECTION OF FISH VIRUSES BY REAL-TIME PCR 审中-公开
公开(公告)号：US20090181363A1
公开(公告)日：2009-07-16
申请号：US12158096
申请日：2006-12-21
申请人： Arun K. Dhar
发明人： Arun K. Dhar
IPC分类号： C12Q1/70 , C12Q1/68
CPC分类号： C12Q1/686 , C12Q2561/113
摘要： A real-time assay coupled with a non-invasive tissue sampling was developed for the detection and quantification of fish viruses. As a proof of principles, data were presented for the detection and quantification of infectious hypodermal necrosis virus (IHNV) in trout. The primers were designed for IHNV nucleocapsid (N), and surface glycoprotein (G) genes, and trout &bgr;-actin and elongation factor-l&agr; (EF-I &agr;) were used as internal control for the assay. The reaction conditions for the real-time RT-PCR were optimized using cDNA derived from IHNV-infected Epithelioma papulosum cyprinid (EPC) cells. Using both N- and G-gene primers, IHNV was successfully detected in liver, kidney, spleen, adipose tissue and pectoral fin samples of laboratory-challenged and wild samples. The dissociation curves with a single melting peak at expected temperature (85° C. for the N-gene and 86.5° C. for the G-gene) confirmed the specificity of the N- and G-gene amplicons. The IHNV N- and the G-gene expression levels in different tissues of laboratory challenged samples were in the order of spleen, liver, kidney, adipose tissue and pectoral fin, however in the field-collected samples the order of gene expression was liver, kidney, pectoral fin, adipose tissue, and spleen. The N- and G-gene expressions in spleen were found to be dramatically lower in the field-collected samples compared to the laboratory-challenged samples indicating a potential difference in the IHNV replication in the laboratory as opposed to field conditions. The real-time PCR assay was found to be rapid, highly sensitive, and reproducible. Based upon the ability to detect the virus in pectoral fins a non-invasive detection method for IHNV and other fish viruses is developed. Such a non-invasive tissue sampling coupled with real-time PCR assay is very valuable for large-scale virus screening of fish in aquaculture facilities as well as for epidemiological studies.

6. 发明授权

US08221767B2 Antigenicity of infectious pancreatic necrosis virus VP2 sub-viral particles expressed in yeast 有权
标题翻译：感染性胰腺坏死病毒VP2亚病毒颗粒在酵母中表达的抗原性
公开(公告)号：US08221767B2
公开(公告)日：2012-07-17
申请号：US12519991
申请日：2007-12-18
申请人： Arun K. Dhar , Robert M. Bowers , F. C. Thomas Allnutt
发明人： Arun K. Dhar , Robert M. Bowers , F. C. Thomas Allnutt
IPC分类号： A61K38/00 , C12P19/34
CPC分类号： A61K39/12 , A61K2039/51 , A61K2039/5258 , A61K2039/542 , A61K2039/552 , C07K14/005 , C12N7/00 , C12N2720/10022 , C12N2720/10023 , C12N2720/10034
摘要： Infectious pancreatic necrosis virus (IPNV), the etiologic agent of infectious pancreatic necrosis in salmonid fish, causes significant losses to the aquaculture industry. The gene for the viral capsid protein (VP2) was cloned into a yeast expression vector and expressed in Saccharomyces cerevisae. Expression of the capsid gene in yeast resulted in formation of approximately 20 nanometer sub-viral particles composed solely of VP2 protein. Anti-IPNV antibodies were detected in rainbow trout vaccinated either by injection of purified VP2-subviral particles (rVP2-SVP) or by feeding recombinant yeast expressing rVP2-SVP. Challenge of rVP2-SVP immunized trout with a heterologous IPNV strain and subsequent viral load determination showed that both injection and orally vaccinated fish had lower IPNV loads than naive or sham-vaccinated fish. This study demonstrates the ability of rVP2-SVPs to induce a specific immune response and the ability of immunized fish to reduce the viral load after an experimentally induced IPNV infection. The invention is not limited to IPNV, and is applicable to other similar viruses for which SVPs can be made and administered to fish.
摘要翻译：传染性胰腺坏死病毒（IPNV）是鲑鱼感染性胰腺坏死的病原体，对水产养殖业造成重大损失。将病毒衣壳蛋白（VP2）的基因克隆到酵母表达载体中并在酿酒酵母中表达。酵母中衣壳基因的表达导致形成仅由VP2蛋白组成的约20纳米亚病毒颗粒。通过注射纯化的VP2-亚病毒颗粒（rVP2-SVP）或通过加入表达rVP2-SVP的重组酵母，在虹鳟中检测到抗IPNV抗体。用异源IPNV毒株对rVP2-SVP免疫的鳟鱼进行挑战，随后的病毒载量测定显示注射和口服接种的鱼类比幼稚或假手术的鱼具有较低的IPNV负荷。该研究证明rVP2-SVP诱导特异性免疫应答的能力以及免疫鱼在实验诱导的IPNV感染后降低病毒载量的能力。本发明不限于IPNV，适用于可以制造和施用SVP的其他类似病毒。

7. 发明授权

US07973148B2 Crustacean expression vector 有权
标题翻译：甲壳动物表达载体
公开(公告)号：US07973148B2
公开(公告)日：2011-07-05
申请号：US11587770
申请日：2005-04-15
申请人： Arun K. Dhar , F. C. Thomas Allnutt
发明人： Arun K. Dhar , F. C. Thomas Allnutt
IPC分类号： C07H21/02 , C07H21/04 , A01N63/00 , A01N65/00 , A61K48/00 , C12N5/00 , C12N5/02
CPC分类号： C12N15/85 , A01K2227/70 , A01K2267/0393 , C12N2799/021 , C12N2800/90
摘要： Methods and constructs for genetic manipulation of one or more of shrimp, shellfish, mollusks, and fish are disclosed. The nucleic acid construct includes a promoter and an internal ribosome entry site of an insect picomavirus, such as a cricket paralysis-like picomavirus. One or more open reading frames can be operably associated with one or both of the promoter and the internal ribosome entry site, and one or more proteins or protein subunits can be expressed upon introduction of the construct into a host cell, such as into a shrimp. Method for producing immortalized crustacean cell lines using enhancer elements derived from shrimp and/or shrimp viruses are also described.
摘要翻译：公开了一种或多种虾，贝类，软体动物和鱼的遗传操作的方法和构造。核酸构建体包括一种启动子和一种诸如板球麻痹状皮莫瓦病毒之类的皮肤微病毒病毒的内部核糖体进入位点。一个或多个开放阅读框可以与启动子和内部核糖体进入位点中的一个或两个可操作地相关联，并且一个或多个蛋白质或蛋白质亚基可以在将构建体引入宿主细胞时表达，例如进入虾。还描述了使用源自虾和/或虾病毒的增强子元件生产永生化甲壳类细胞系的方法。

8. 发明申请

US20130195914A1 Expression Of Positive Sense Single Stranded RNA Virus And Uses Thereof 审中-公开
标题翻译：阳性单链RNA病毒的表达及其用途
公开(公告)号：US20130195914A1
公开(公告)日：2013-08-01
申请号：US13640625
申请日：2011-04-08
申请人： Arun K. Dhar
发明人： Arun K. Dhar
IPC分类号： C12N15/866 , A61K39/29 , A61K39/12 , C12N15/64
CPC分类号： C12N15/866 , A61K39/12 , A61K39/29 , A61K2039/5252 , C12N7/00 , C12N15/64 , C12N15/86 , C12N2710/14143 , C12N2710/14144 , C12N2770/10052 , C12N2770/22052 , C12N2770/24152 , C12N2770/24234 , C12N2770/24252 , Y02A50/394
摘要： The invention relates to the fields of viruses, vaccines and compounds and methods for expression. In particular, the invention includes methods and agents capable of producing quantities of a vaccine to a positive sense single stranded RNA (“(+)sense RNA”)virus.
摘要翻译：本发明涉及病毒，疫苗和化合物的领域以及用于表达的方法。特别地，本发明包括能够产生大量针对正义单链RNA（“（+）有义RNA”）病毒的疫苗的方法和试剂。

9. 发明授权

US09345760B2 IPNV-ISAV bivalent vaccine using a virus-like particle-based platform and methods of using the same 有权
标题翻译： IPNV-ISAV二价疫苗使用基于病毒的粒子平台及其使用方法
公开(公告)号：US09345760B2
公开(公告)日：2016-05-24
申请号：US14343666
申请日：2012-09-07
申请人： Arun K. Dhar
发明人： Arun K. Dhar
IPC分类号： C07K7/06 , A61K39/295 , A61K39/12 , C07K5/103 , C12N7/00 , A61K39/00
CPC分类号： A61K39/295 , A61K39/12 , A61K2039/523 , A61K2039/542 , A61K2039/552 , A61K2039/55566 , A61K2039/70 , C07K5/101 , C07K7/06 , C07K2319/00 , C12N7/00 , C12N2720/10023 , C12N2720/10034 , C12N2720/10044 , C12N2720/12023 , C12N2720/12034 , C12N2760/16034 , C12N2760/16043
摘要： The present invention provides bivalent oral vaccines against infectious pancreatic necrosis virus (IPNV) and infectious salmon anemia virus (ISAV). Yeast cells comprise an expression vector comprising (i) a polynucleotide sequence encoding a VP2 capsid protein of IPNV and (ii) a polynucleotide sequence encoding one or more antigenic epitopes of hemaglutinin of ISAV. The yeast cells express sub-viral particles (SVP) comprising a VP2 capsid protein of IPNV and one or more antigenic epitopes of hemaglutinin of ISAV. The yeast cells and SVP may be administered in an effective amount to increase the amount of antibodies against IPNV and ISAV in the fish, preferably Salmonidae. The yeast cells and SVP may be in the form of fish food for administering to the fish orally.
摘要翻译：本发明提供针对感染性胰腺坏死病毒（IPNV）和感染性鲑鱼贫血病毒（ISAV）的二价口服疫苗。酵母细胞包含表达载体，其包含（i）编码IPNV的VP2衣壳蛋白的多核苷酸序列和（ii）编码ISAV血凝素的一个或多个抗原表位的多核苷酸序列。酵母细胞表达包含IPNV的VP2衣壳蛋白和ISAV的血凝素的一个或多个抗原表位的亚病毒颗粒（SVP）。可以以有效量施用酵母细胞和SVP以增加鱼，优选鲑科的IPNV和ISAV抗体的量。酵母细胞和SVP可以是鱼类的形式，用于口服给鱼。

10. 发明申请

US20080194504A1 Rna-Mediated Interference to Control Disease in Terrestrial and Aquaculture Animals 审中-公开
标题翻译： Rna介导的对陆地和水产养殖动物控制疾病的干扰
公开(公告)号：US20080194504A1
公开(公告)日：2008-08-14
申请号：US10588414
申请日：2005-02-04
申请人： David J. Kyle , Arun K. Dhar
发明人： David J. Kyle , Arun K. Dhar
IPC分类号： A61K31/7105 , A61P31/04 , A61P31/10 , A61P31/12
CPC分类号： C12N15/111 , C12N15/1131 , C12N2310/111 , C12N2310/14 , C12N2310/53 , C12N2320/32
摘要： The invention is directed to compositions (e.g., feeds, feed supplements, and therapeutic products) and methods for inhibiting an animal pathogen using RNA-interference technology.
摘要翻译：本发明涉及使用RNA干扰技术抑制动物病原体的组合物（例如饲料，饲料添加剂和治疗产品）和方法。

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