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1. 发明申请

WO2006019876A3 PRODUCTION OF FUSION PROTEINS BY CELL-FREE PROTEIN SYNTHESIS 审中-公开
标题翻译：通过无细胞蛋白合成生产融合蛋白
公开(公告)号：WO2006019876A3
公开(公告)日：2009-04-16
申请号：PCT/US2005024940
申请日：2005-07-14
申请人： INVITROGEN CORP , KUDLICKI WIESLAW ANTONI , FLETCHER JULIA , KATZEN FEDERICO , BENNET ROBERT
发明人： KUDLICKI WIESLAW ANTONI , FLETCHER JULIA , KATZEN FEDERICO , BENNET ROBERT
IPC分类号： C07H21/04 , C12N9/98 , C12P21/06
CPC分类号： C12P21/02
摘要： The present invention relates to in vitro protein synthesis (IVPS) systems, particularly such systems using suppressor tRNAs and rare codon tRNAs to extend translation of an open reading frame into fusion protein elements, thereby generating fusion proteins in vitro. The invention also provides methods, compositions and kits using the IVPS systems of the invention, and proteins produced using the methods, compositions, kits and IVPS systems of the invention.
摘要翻译：本发明涉及体外蛋白质合成（IVPS）系统，特别是使用抑制子tRNA和稀有密码子tRNA的系统将开放阅读框的翻译扩展到融合蛋白质元件中，从而在体外产生融合蛋白。本发明还提供使用本发明的IVPS系统的方法，组合物和试剂盒，以及使用本发明的方法，组合物，试剂盒和IVPS系统产生的蛋白质。

2. 发明申请

WO2008106660A3 ISOLATED PHOSPHOLIPID-PROTEIN PARTICLES 审中-公开
标题翻译：分离的磷脂 - 蛋白质颗粒
公开(公告)号：WO2008106660A3
公开(公告)日：2008-11-20
申请号：PCT/US2008055567
申请日：2008-02-29
申请人： INVITROGEN CORP , KATZEN FEDERICO , FLETCHER JULIA , KUDLICKI WIESLAW , BEECHEM JOSEPH , WANG LILIN
发明人： KATZEN FEDERICO , FLETCHER JULIA , KUDLICKI WIESLAW , BEECHEM JOSEPH , WANG LILIN
IPC分类号： C07K14/47 , C07K1/00 , C12P21/06 , C12Q1/68
CPC分类号： C12P21/02 , C07K1/02
摘要： Systems and methods are provided for producing a protein of interest that is typically not amenable to expression in soluble form in in vitro expression systems. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include a scaffold protein such as apolipoprotein or an amphipathic alpha helix containing ("AAHC") protein, in which higher yields of soluble protein are produced than in the absence of the scaffold protein. The scaffold proteins may be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The scaffold protein may be provided as a protein per se or may be encoded by a nucleic acid template and co-expressed with the protein of interest. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein expression and isolation.
摘要翻译：提供了用于产生通常不适于在体外表达系统中以可溶形式表达的目的蛋白质的系统和方法。在一些方面，本发明提供了使用体外蛋白质合成系统合成蛋白质的方法，所述体外蛋白质合成系统包括支架蛋白质例如载脂蛋白或含有（“AAHC”）蛋白质的两亲性α螺旋，其中产生的可溶性蛋白质产量高于缺少支架蛋白。可以在与脂质相关的体外蛋白质合成系统中提供支架蛋白质，或者不提供与脂质相关的支架蛋白质。支架蛋白本身可以作为蛋白质提供，或者可以由核酸模板编码并与感兴趣的蛋白质共表达。本发明还提供用于合成可溶形式的蛋白质的组合物和试剂盒，其中组合物和试剂盒包括用于蛋白质表达和分离的细胞提取物。

3. 发明申请

WO2006078821A3 PRODUCTS AND PROCESSES FOR IN VITRO SYNTHESIS OF BIOMOLECULES 审中-公开
标题翻译：生物分子的体外合成的产物和方法
公开(公告)号：WO2006078821A3
公开(公告)日：2007-02-15
申请号：PCT/US2006001901
申请日：2006-01-20
申请人： INVITROGEN CORP , KATZEN FEDERICO , KUDLICKI WIESLAW , FLETCHER JULIA
发明人： KATZEN FEDERICO , KUDLICKI WIESLAW , FLETCHER JULIA
IPC分类号： C12P21/06 , C12P19/34 , C12P19/40
CPC分类号： C12P19/34 , C12P21/005
摘要： Provided herein are products and processes for efficiently synthesizing biomolecules in vitro using cell-free extracts derived from mammalian cells and insect cells. In an embodiment, provided is a process for preparing a cell-free extract from insect cells and mammalian cells that efficiently synthesizes post-translationally modified target proteins (e.g., glycosylated target proteins). In some embodiments, a ribonucleic acid is synthesized in vitro that comprises a cap, a 5' untranslated region comprising an 18S rRNA binding ribonucleotide sequence, and a target ribonucleotide sequence. It has been determined that such ribonucleic acids result in efficient in vitro synthesis of a target protein using cell-free extracts derived from non-rabbit mammalian cells and insect cells.
摘要翻译：本文提供了使用来自哺乳动物细胞和昆虫细胞的无细胞提取物在体外有效合成生物分子的产物和方法。在一个实施方案中，提供了从昆虫细胞和哺乳动物细胞制备无细胞提取物的方法，其有效合成翻译后修饰的靶蛋白（例如糖基化靶蛋白）。在一些实施方案中，在体外合成核糖核酸，其包含帽，包含18S rRNA结合核糖核苷酸序列的5'非翻译区和靶核糖核苷酸序列。已经确定，使用来自非兔哺乳动物细胞和昆虫细胞的无细胞提取物，这种核糖核酸导致靶蛋白的有效体外合成。

4. 发明申请

WO2008106660A2 ISOLATED PHOSPHOLIPID-PROTEIN PARTICLES 审中-公开
标题翻译：分离的磷脂 - 蛋白质颗粒
公开(公告)号：WO2008106660A2
公开(公告)日：2008-09-04
申请号：PCT/US2008/055567
申请日：2008-02-29
申请人： INVITROGEN CORPORATION , KATZEN, Federico , FLETCHER, Julia , KUDLICKI, Wieslaw , BEECHEM, Joseph , WANG, Lilin
发明人： KATZEN, Federico , FLETCHER, Julia , KUDLICKI, Wieslaw , BEECHEM, Joseph , WANG, Lilin
IPC分类号： C07K14/47 , C07K1/00 , C12Q1/68 , C12P21/06
CPC分类号： C12P21/02 , C07K1/02
摘要： Systems and methods are provided for producing a protein of interest that is typically not amenable to expression in soluble form in in vitro expression systems. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include a scaffold protein such as apolipoprotein or an amphipathic alpha helix containing ("AAHC") protein, in which higher yields of soluble protein are produced than in the absence of the scaffold protein. The scaffold proteins may be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The scaffold protein may be provided as a protein per se or may be encoded by a nucleic acid template and co-expressed with the protein of interest. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein expression and isolation.
摘要翻译：提供的系统和方法用于产生通常不适于在体外表达系统中以可溶形式表达的目标蛋白质。在一些方面，本发明提供使用体外蛋白质合成系统合成蛋白质的方法，所述体外蛋白质合成系统包括支架蛋白如载脂蛋白或含有（“AAHC”）蛋白的两亲性α螺旋，其中产生比在没有支架蛋白。支架蛋白质可以提供在与脂质相关的体外蛋白质合成系统中或与脂质不相关。支架蛋白质可以作为蛋白质本身提供，或者可以由核酸模板编码并与感兴趣的蛋白质共表达。本发明还提供用于合成可溶形式的蛋白质的组合物和试剂盒，其中组合物和试剂盒包括用于蛋白质表达和分离的细胞提取物。

5. 发明申请

WO2006078821A2 PRODUCTS AND PROCESSES FOR IN VITRO SYNTHESIS OF BIOMOLECULES 审中-公开
标题翻译：生物分子的体外合成的产物和方法
公开(公告)号：WO2006078821A2
公开(公告)日：2006-07-27
申请号：PCT/US2006/001901
申请日：2006-01-20
申请人： INVITROGEN CORPORATION , KATZEN, Federico , KUDLICKI, Wieslaw , FLETCHER, Julia
发明人： KATZEN, Federico , KUDLICKI, Wieslaw , FLETCHER, Julia
IPC分类号： C12P21/06
CPC分类号： C12P19/34 , C12P21/005
摘要： Provided herein are products and processes for efficiently synthesizing biomolecules in vitro using cell-free extracts derived from mammalian cells and insect cells. In an embodiment, provided is a process for preparing a cell-free extract from insect cells and mammalian cells that efficiently synthesizes post-translationally modified target proteins (e.g., glycosylated target proteins). In some embodiments, a ribonucleic acid is synthesized in vitro that comprises a cap, a 5' untranslated region comprising an 18S rRNA binding ribonucleotide sequence, and a target ribonucleotide sequence. It has been determined that such ribonucleic acids result in efficient in vitro synthesis of a target protein using cell-free extracts derived from non-rabbit mammalian cells and insect cells.
摘要翻译：本文提供了使用来自哺乳动物细胞和昆虫细胞的无细胞提取物在体外有效合成生物分子的产物和方法。在一个实施方案中，提供了从昆虫细胞和哺乳动物细胞制备无细胞提取物的方法，其有效合成翻译后修饰的靶蛋白（例如糖基化靶蛋白）。在一些实施方案中，在体外合成核糖核酸，其包含帽，包含18S rRNA结合核糖核苷酸序列的5'非翻译区和靶核糖核苷酸序列。已经确定，使用来自非兔哺乳动物细胞和昆虫细胞的无细胞提取物，这种核糖核酸导致靶蛋白的有效体外合成。

6. 发明申请

WO2006039622A3 FEEDING BUFFERS, SYSTEMS, AND METHODS FOR IN VITRO SYNTHESIS OF BIOMOLECULES 审中-公开
公开(公告)号：WO2006039622A3
公开(公告)日：2006-04-13
申请号：PCT/US2005/035419
申请日：2005-10-03
申请人： INVITROGEN CORPORATION , KUDLICKI, Wieslaw, Antoni , KEPPETIPOLA, Shiranthi , FLETCHER, Julia , GETBEHEAD, Ashley Elaine , KATZEN, Federico , VOZZA-BROWN, Laura
发明人： KUDLICKI, Wieslaw, Antoni , KEPPETIPOLA, Shiranthi , FLETCHER, Julia , GETBEHEAD, Ashley Elaine , KATZEN, Federico , VOZZA-BROWN, Laura
IPC分类号： C12Q1/68 , C12P21/06 , C07K1/00
摘要： Compositions, methods and kits for in vitro systems for synthesis of biomolecules such as polypeptides, are provided herein. Cell extracts that provide enhanced yields of soluble proteins using in vitro protein synthesis methods are provided. The invention also includes methods for producing high yields of proteins by the addition of a feeding solution that includes amino acids and an energy source to an ongoing in vitro synthesis system. The invention also includes methods of using a high-yield in vitro synthesis system to produce large quantities of proteins with incorporated labeled amino acids for analysis by methods such as by NMR. The invention further includes vectors for enhanced production of proteins from nucleic acid templates using in vitro synthesis systems.

7. 发明申请

WO2007038755A1 IN VITRO PROTEIN SYNTHESIS SYSTEMS FOR MEMBRANE PROTEINS THAT INCLUDE APOLIPOPROTEINS AND PHOSPHOLIPID-APOLIPOPROTEIN PARTICLES 审中-公开
标题翻译：用于包含APOLIPOPROTEINS和磷脂 - 磷脂酰胺颗粒的膜蛋白的体外蛋白合成系统
公开(公告)号：WO2007038755A1
公开(公告)日：2007-04-05
申请号：PCT/US2006/038173
申请日：2006-09-27
申请人： INVITROGEN CORPORATION , KUDLICKI, Wieslaw , FLETCHER, Julia , KATZEN, Federico
发明人： KUDLICKI, Wieslaw , FLETCHER, Julia , KATZEN, Federico
IPC分类号： C12P21/02 , C12N1/00
CPC分类号： C12P21/02 , C07K14/775
摘要： In vitro protein synthesis systems and methods are provided that produce membrane proteins in soluble form. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include an apolipoprotein, in which higher yields of soluble protein are produced than in the absence of the apolipoprotein. Apolipoproteins useful in the present invention include naturally occurring apolipoproteins, as well as sequence variants of wild-type apolipoproteins, and engineered apolipoproteins. The apolipoproteins can be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein translation and at least one apolipoprotein biomolecule.
摘要翻译：提供了产生可溶形式的膜蛋白的体外蛋白质合成系统和方法。在一些方面，本发明提供了使用体外蛋白质合成系统合成蛋白质的方法，所述体外蛋白质合成系统包括与不存在载脂蛋白相比产生更高产率的可溶性蛋白质的载脂蛋白。可用于本发明的载脂蛋白包括天然存在的载脂蛋白，以及野生型载脂蛋白和工程化载脂蛋白的序列变体。载脂蛋白可以在与脂质相关的体外蛋白质合成系统中提供，或与脂质不相关。本发明还提供用于合成可溶形式的蛋白质的组合物和试剂盒，其中组合物和试剂盒包括用于蛋白质翻译的细胞提取物和至少一种载脂蛋白生物分子。

8. 发明申请

WO2006039622A2 FEEDING BUFFERS, SYSTEMS, AND METHODS FOR IN VITRO SYNTHESIS OF BIOMOLECULES 审中-公开
标题翻译：饲料缓冲液，系统和方法用于生物分子的体外合成
公开(公告)号：WO2006039622A2
公开(公告)日：2006-04-13
申请号：PCT/US2005035419
申请日：2005-10-03
申请人： INVITROGEN CORP , KUDLICKI WIESLAW ANTONI , KEPPETIPOLA SHIRANTHI , FLETCHER JULIA , GETBEHEAD ASHLEY ELAINE , KATZEN FEDERICO , VOZZA-BROWN LAURA
发明人： KUDLICKI WIESLAW ANTONI , KEPPETIPOLA SHIRANTHI , FLETCHER JULIA , GETBEHEAD ASHLEY ELAINE , KATZEN FEDERICO , VOZZA-BROWN LAURA
IPC分类号： C12P21/06 , C12N15/74
CPC分类号： C12P21/00 , C07K1/02 , C12N15/70 , C12P21/02
摘要： Compositions, methods and kits for in vitro systems for synthesis of biomolecules such as polypeptides, are provided herein. Cell extracts that provide enhanced yields of soluble proteins using in vitro protein synthesis methods are provided. The invention also includes methods for producing high yields of proteins by the addition of a feeding solution that includes amino acids and an energy source to an ongoing in vitro synthesis system. The invention also includes methods of using a high-yield in vitro synthesis system to produce large quantities of proteins with incorporated labeled amino acids for analysis by methods such as by NMR. The invention further includes vectors for enhanced production of proteins from nucleic acid templates using in vitro synthesis systems.
摘要翻译：本文提供了用于合成生物分子如多肽的体外系统的组合物，方法和试剂盒。提供了使用体外蛋白质合成方法提供增加的可溶性蛋白质产量的细胞提取物。本发明还包括通过添加包括氨基酸和进入体外合成系统的能量源的进料溶液来生产高产量蛋白质的方法。本发明还包括使用高产量体外合成系统产生大量具有掺入标记氨基酸的蛋白质以通过诸如NMR的方法进行分析的方法。本发明还包括使用体外合成系统从核酸模板增强蛋白质生产的载体。

9. 发明申请

WO2006019876A2 PRODUCTION OF FUSION PROTEINS BY CELL-FREE PROTEIN SYNTHESIS 审中-公开
标题翻译：通过无细胞蛋白合成生产融合蛋白
公开(公告)号：WO2006019876A2
公开(公告)日：2006-02-23
申请号：PCT/US2005/024940
申请日：2005-07-14
申请人： INVITROGEN CORPORATION , KUDLICKI, Wieslaw, Antoni , FLETCHER, Julia , KATZEN, Federico , BENNET, Robert
发明人： KUDLICKI, Wieslaw, Antoni , FLETCHER, Julia , KATZEN, Federico , BENNET, Robert
IPC分类号： C12P21/06
CPC分类号： C12P21/02
摘要： The present invention relates to in vitro protein synthesis (IVPS) systems, particularly such systems using suppressor tRNAs and rare codon tRNAs to extend translation of an open reading frame into fusion protein elements, thereby generating fusion proteins in vitro. The invention also provides methods, compositions and kits using the IVPS systems of the invention, and proteins produced using the methods, compositions, kits and IVPS systems of the invention.
摘要翻译：本发明涉及体外蛋白质合成（IVPS）系统，特别是使用抑制子tRNA和稀有密码子tRNA的系统将开放阅读框的翻译扩展到融合蛋白质元件中，从而在体外产生融合蛋白。本发明还提供使用本发明的IVPS系统的方法，组合物和试剂盒，以及使用本发明的方法，组合物，试剂盒和IVPS系统产生的蛋白质。

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