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1. 发明授权

US07939710B1 Trophic conversion of obligate phototrophic algae through metabolic engineering 失效
公开(公告)号：US07939710B1
公开(公告)日：2011-05-10
申请号：US09839536
申请日：2001-04-23
申请人： Kirk Emil Apt , F. C. Thomas Allnutt , David J. Kyle , James Casey Lippmeier
发明人： Kirk Emil Apt , F. C. Thomas Allnutt , David J. Kyle , James Casey Lippmeier
IPC分类号： C12N15/82
CPC分类号： C12N15/8209 , C12N1/12 , C12N15/65 , C12N15/79 , C12N15/8207 , Y10S435/946
摘要： 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.

2. 发明授权

US08008061B2 Trophic conversion of obligate phototrophic algae through metabolic engineering 失效
标题翻译：通过代谢工程营养转化专性光营养藻类
公开(公告)号：US08008061B2
公开(公告)日：2011-08-30
申请号：US11842898
申请日：2007-08-21
申请人： Kirk Emil Apt , F. C. Thomas Allnutt , David J. Kyle , James Casy Lippmeier
发明人： Kirk Emil Apt , F. C. Thomas Allnutt , David J. Kyle , James Casy Lippmeier
IPC分类号： C12N1/12
CPC分类号： C12N15/8209 , C12N1/12 , C12N15/65 , C12N15/79 , C12N15/8207 , Y10S435/946
摘要： 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.
摘要翻译：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利性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。

3. 发明授权

US6027900A Methods and tools for transformation of eukaryotic algae 有权
标题翻译：用于转化真核藻类的方法和工具
公开(公告)号：US6027900A
公开(公告)日：2000-02-22
申请号：US155972
申请日：1998-10-09
申请人： F. C. Thomas Allnutt , David J. Kyle , Arthur R. Grossman , Kirk R. Apt
发明人： F. C. Thomas Allnutt , David J. Kyle , Arthur R. Grossman , Kirk R. Apt
IPC分类号： C12N15/65 , C12N15/79 , C12Q1/68
CPC分类号： C12N15/79 , C12N15/65
摘要： Genetic fusions for use in genetic engineering of eukaryotic algae employ a promoter from a light harvesting protein fused to a protein of interest. The fusions can be introduced and selected using an antibiotic resistance determinant. One gene useful for such selection is the sh ble gene encoding a bleomycin binding protein.
摘要翻译： PCT No.PCT / US97 / 06021 Sec。 371日期：1998年10月9日 102（e）日期1998年10月9日PCT 1997年4月11日PCT PCT。 WO97 / 39106 PCT公开号日期1997年10月23日用于真核藻类遗传工程的遗传融合使用与感兴趣的蛋白质融合的光收获蛋白质的启动子。可以使用抗生素抗性决定簇引入和选择融合体。用于这种选择的一个基因是编码博来霉素结合蛋白的sh ble基因。

4. 发明授权

US07550647B2 Transfected shrimp as production systems for therapeutic proteins 有权
标题翻译：转染的虾作为治疗性蛋白质的生产系统
公开(公告)号：US07550647B2
公开(公告)日：2009-06-23
申请号：US10778175
申请日：2004-02-17
申请人： Ruth Barratt , F. C. Thomas Allnutt , Robert Bullis , David J. Kyle
发明人： Ruth Barratt , F. C. Thomas Allnutt , Robert Bullis , David J. Kyle
IPC分类号： A01K67/00 , A01K67/033
CPC分类号： C07K14/005 , A01K2227/70 , A01K2267/01 , A61K2039/5256 , C07K14/505 , C07K14/62 , C07K16/00 , C07K2317/10 , C12N7/00 , C12N9/16 , C12N15/86 , C12N2710/14043 , C12N2710/18043 , C12N2750/14043 , C12N2770/16022 , C12N2770/16034 , C12N2770/22043 , C12N2770/36143 , C12N2830/00 , C12N2830/75
摘要： A crustacean or rotifer is infected with a recombinant infectious virus that expresses a protein exogenous to the virus. The genome of the crustacean or rotifer itself remains unaltered. Crustacean, rotifer, insect, or viral promoters drive the transcription of a gene inserted into the recombinant virus genome, and the virus replicates in the crustacean or rotifer cell cytoplasm. The infected crustacean or rotifer can be provided directly to humans or non-human animals, or, following production and harvest of the crustaceans or rotifers, purified recombinant protein or polypeptide can be provided. Large quantities of biopharmaceuticals can be produced rapidly and inexpensively using this production system.
摘要翻译：用表达外源蛋白质的病毒的重组感染性病毒感染甲壳类动物或轮虫。甲壳类或轮虫本身的基因组本身保持不变。甲壳动物，轮虫，昆虫或病毒启动子驱动插入重组病毒基因组中的基因的转录，并且病毒在甲壳类或轮状细胞细胞质中复制。受感染的甲壳动物或轮虫可以直接提供给人类或非人动物，或者在甲壳类动物或轮虫的生产和收获后，可提供纯化的重组蛋白或多肽。可以使用该生产系统快速且廉价地生产大量的生物药物。

5. 发明授权

US07932056B2 Crustaceans as production systems for therapeutic proteins 有权
标题翻译：甲壳动物作为治疗性蛋白质的生产系统
公开(公告)号：US07932056B2
公开(公告)日：2011-04-26
申请号：US12489905
申请日：2009-06-23
申请人： Ruth Barratt , F. C. Thomas Allnutt , Robert Bullis , David J. Kyle
发明人： Ruth Barratt , F. C. Thomas Allnutt , Robert Bullis , David J. Kyle
IPC分类号： C12P21/00 , C12N5/06 , C12N7/00
CPC分类号： C07K14/005 , A01K2227/70 , A01K2267/01 , A61K2039/5256 , C07K14/505 , C07K14/62 , C07K16/00 , C07K2317/10 , C12N7/00 , C12N9/16 , C12N15/86 , C12N2710/14043 , C12N2710/18043 , C12N2750/14043 , C12N2770/16022 , C12N2770/16034 , C12N2770/22043 , C12N2770/36143 , C12N2830/00 , C12N2830/75
摘要： A crustacean or rotifer is infected with a recombinant infectious virus that expresses a protein exogenous to the virus. The genome of the crustacean or rotifer itself remains unaltered. Crustacean, rotifer, insect, or viral promoters drive the transcription of a gene inserted into the recombinant virus genome, and the virus replicates in the crustacean or rotifer cell cytoplasm. The infected crustacean or rotifer can be provided directly to humans or non-human animals, or, following production and harvest of the crustaceans or rotifers, purified recombinant protein or polypeptide can be provided. Large quantities of biopharmaceuticals can be produced rapidly and inexpensively using this production system.
摘要翻译：用表达外源蛋白质的病毒的重组感染性病毒感染甲壳类动物或轮虫。甲壳类或轮虫本身的基因组本身保持不变。甲壳动物，轮虫，昆虫或病毒启动子驱动插入重组病毒基因组中的基因的转录，并且病毒在甲壳类或轮状细胞细胞质中复制。受感染的甲壳动物或轮虫可以直接提供给人类或非人动物，或者在甲壳类动物或轮虫的生产和收获后，可提供纯化的重组蛋白或多肽。可以使用该生产系统快速且廉价地生产大量的生物药物。

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. 发明授权

US07256050B2 High fluorescent intensity cross-linked allophycocyanin 失效
标题翻译：高荧光强度交联别藻蓝蛋白
公开(公告)号：US07256050B2
公开(公告)日：2007-08-14
申请号：US09882376
申请日：2001-06-18
申请人： John Peter Morseman , Mark Wesley Moss , F. C. Thomas Allnutt
发明人： John Peter Morseman , Mark Wesley Moss , F. C. Thomas Allnutt
IPC分类号： G01N21/76
CPC分类号： G01N33/582 , G01N33/533 , G01N33/542
摘要： In a method for quantitating an analyte by measuring time resolved transfer of fluorescence energy to or from a label quantitatively associated with the analyte, the present invention provides an improvement comprising measuring the energy transferred from donor compounds having the ability to absorb light energy and then transfer this energy to cross-linked allophycocyanin in a time-resolved manner, where the cross-linked allophycocyanin used according to this invention has not been exposed to strongly chaotropic agents after cross-linking.
摘要翻译：在通过测量与分析物定量相关的标记物的荧光能量的时间分辨转移的方法来定量分析物的方法，本发明提供了一种改进，其包括测量从具有吸收光能并且然后转移的能力的给体化合物转移的能量该能量以时间分辨的方式交联别藻蓝蛋白，其中根据本发明使用的交联别藻蓝蛋白在交联后未暴露于强离液剂。

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