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    • 1. 发明申请
    • CRUSTACEAN EXPRESSION VECTOR
    • 表达式表达向量
    • US20110269225A1
    • 2011-11-03
    • US13176026
    • 2011-07-05
    • ARUN K. DHARF.C. THOMAS ALLNUTT
    • ARUN K. DHARF.C. THOMAS ALLNUTT
    • C12N15/85
    • C12N15/85A01K2227/70A01K2267/0393C12N2799/021C12N2800/90
    • 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.
    • 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。
    • 4. 发明申请
    • ANTIGENICITY OF INFECTIOUS PANCREATIC NECROSIS VIRUS VP2 SUB-VIRAL PARTICLES EXPRESSED IN YEAST
    • 感染性胰腺炎病毒VP2亚历山大小肠病毒的抗生素
    • US20100092521A1
    • 2010-04-15
    • US12519991
    • 2007-12-18
    • Arun K. DharRobert M. BowersF.C. Thomas Allnutt
    • Arun K. DharRobert M. BowersF.C. Thomas Allnutt
    • A61K39/00C12N1/19C12P21/00A61P31/12
    • A61K39/12A61K2039/51A61K2039/5258A61K2039/542A61K2039/552C07K14/005C12N7/00C12N2720/10022C12N2720/10023C12N2720/10034
    • 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.
    • 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。
    • 5. 发明申请
    • TROPHIC CONVERSION OF OBLIGATE PHOTOTROPHIC ALGAE THROUGH METABOLIC ENGINEERING
    • 通过代谢工程进行的白藜芦醇的热转化
    • US20080138851A1
    • 2008-06-12
    • US11842898
    • 2007-08-21
    • Kirk Emil AptF.C. Thomas AllnuttDavid J. KyleJames Casey Lippmeier
    • Kirk Emil AptF.C. Thomas AllnuttDavid J. KyleJames Casey Lippmeier
    • C12Q1/02C12N1/12C12N15/74
    • C12N15/8209C12N1/12C12N15/65C12N15/79C12N15/8207Y10S435/946
    • Most microalgae are obligate photoautotrophs and their growth is strictly dependent on the generation of photosynthetically-derived energy. In this study it is shown that the microalga Phaeodaclylurn tricornutum can be engineered to import glucose and grow in the dark through the introduction of genes encoding glucose transporters. Both the human and Chlorella kessleri glucose transporters facilitated the uptake of glucose by P. tricornutum, allowing the cells to metabolize exogenous organic carbon and thrive, independent of light. This is the first successful trophic conversion of an obligate photoautotroph through metabolic engineering, and it demonstrates that methods of cell nourishment can be fundamentally altered with the introduction of a single gene. Since strains transformed with the glucose transport genes are able to grow non-photosynthetically, they can be exploited for the analysis of photosynthetic processes through mutant generation and characterization. Finally, this work also represents critical progress toward large-scale commercial exploitation of obligate phototrophic algae through the use of microbial fermentation technology, eliminating significant limitations resulting from light-dependent growth.
    • 大多数微藻是专性光自养体,其生长严格依赖于光合作用衍生能量的产生。 在这项研究中,显示微藻Phaeodaclylurn三角褐豆可以通过引入编码葡萄糖转运蛋白的基因进行工程化以导入葡萄糖并在黑暗中生长。 人和小球藻凯斯勒葡萄糖转运蛋白都促进了三角褐指藻的葡萄糖摄取,使得细胞代谢外源有机碳,并且独立于光。 这是通过代谢工程首次成功地营养专一性自养型营养转化,并且表明通过引入单一基因可以从根本上改变细胞营养的方法。 由于用葡萄糖转运基因转化的菌株能够非光合作用生长,因此可以通过突变体的产生和表征来开发光合作用的分析。 最后,这项工作也是通过使用微生物发酵技术大规模商业开发专用光营养藻类的重要进展,消除了光依赖性生长造成的重大限制。
    • 7. 发明申请
    • Trophic Conversion of Obligate Phototrophic Algae Through Metabolic Engineering
    • 通过代谢工程营养转化特种光营养藻类
    • US20120034653A1
    • 2012-02-09
    • US13188806
    • 2011-07-22
    • Kirk Emil APTF.C. Thomas AllnuttDavid J. KyleJames Casey Lippmeier
    • Kirk Emil APTF.C. Thomas AllnuttDavid J. KyleJames Casey Lippmeier
    • C12P21/00C12N15/79C12N1/13C07C29/74C11B1/10C07C35/21C07C7/00C07K14/405A23K1/00C07K1/14
    • C12N15/8209C12N1/12C12N15/65C12N15/79C12N15/8207Y10S435/946
    • Most microalgae are obligate photoautotrophs and their growth is strictly dependent on the generation of photosynthetically-derived energy. In this study it is shown that the microalga Phaeodaclylurn tricornutum can be engineered to import glucose and grow in the dark through the introduction of genes encoding glucose transporters. Both the human and Chlorella kessleri glucose transporters facilitated the uptake of glucose by P. tricornutum, allowing the cells to metabolize exogenous organic carbon and thrive, independent of light. This is the first successful trophic conversion of an obligate photoautotroph through metabolic engineering, and it demonstrates that methods of cell nourishment can be fundamentally altered with the introduction of a single gene. Since strains transformed with the glucose transport genes are able to grow non-photosynthetically, they can be exploited for the analysis of photosynthetic processes through mutant generation and characterization. Finally, this work also represents critical progress toward large-scale commercial exploitation of obligate phototrophic algae through the use of microbial fermentation technology, eliminating significant limitations resulting from light-dependent growth.
    • 大多数微藻是专性光自养体,其生长严格依赖于光合作用衍生能量的产生。 在这项研究中,显示微藻Phaeodaclylurn三角褐豆可以通过引入编码葡萄糖转运蛋白的基因进行工程化以导入葡萄糖并在黑暗中生长。 人和小球藻凯斯勒葡萄糖转运蛋白都促进了三角褐指藻的葡萄糖摄取,使得细胞代谢外源有机碳,并且独立于光。 这是通过代谢工程首次成功地营养专一性自养型营养转化,并且表明通过引入单一基因可以从根本上改变细胞营养的方法。 由于用葡萄糖转运基因转化的菌株能够非光合作用生长,因此可以通过突变体的产生和表征来开发光合作用的分析。 最后,这项工作也是通过使用微生物发酵技术大规模商业开发专用光营养藻类的重要进展,消除了光依赖性生长造成的重大限制。