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2. 发明申请

US20110150861A1 SORF CONSTRUCTS AND MULTIPLE GENE EXPRESSION 审中-公开
标题翻译： SORF构造和多基因表达
公开(公告)号：US20110150861A1
公开(公告)日：2011-06-23
申请号：US12914556
申请日：2010-10-28
申请人： Gerald R. Carson , Wendy R. Gion , Yune Z. Kunes , Walter F. Leise, III , Rachel A. Davis-Taber , Emma Fung
发明人： Gerald R. Carson , Wendy R. Gion , Yune Z. Kunes , Walter F. Leise, III , Rachel A. Davis-Taber , Emma Fung
IPC分类号： A61K39/395 , C12N15/63 , C12N1/00 , C12N1/21 , C12N1/11 , C12N5/10 , C12N1/19 , C12N1/15 , C12P21/00 , C07K16/00 , C07K16/24 , A61K38/00 , A61P43/00
CPC分类号： C07K16/00 , C07K16/241 , C07K16/244 , C07K2317/14 , C07K2317/21 , C07K2317/76 , C07K2319/50 , C07K2319/92
摘要： Embodiments of the invention relate to vector constructs and methods for expression of polypeptides including multimeric products such as therapeutic antibodies. Particular constructs allow for the generation of expression products from a single open reading frame (sORF). An embodiment provides an isolated or purified expression vector for generating one or more recombinant protein products comprising a single open reading frame insert; said insert comprising a signal peptide nucleic acid sequence encoding a signal peptide; a first nucleic acid sequence encoding a first polypeptide; a first intervening nucleic acid sequence encoding a first protein cleavage site, wherein said first protein cleavage site is provided by an intein segment of a Ion protease gene of Pyrococcus or a klbA gene of Pyrococcus or Methanococcus, or a modified intein segment derived therefrom; and a second nucleic acid sequence encoding a second polypeptide. Certain embodiments of constructs and methods employ an intein segment of a Ion protease gene of Pyrococcus abyssi, Pyrococcus furiosus, or Pyrococcus horikoshii OT3; or an intein segment of a klbA gene of Pyrococcus abyssi, Pyrococcus furiosus, or Methanococcus jannaschii; or other intein segment.
摘要翻译：本发明的实施方案涉及用于表达多肽的载体构建体和方法，包括多聚体产物，例如治疗性抗体。特定结构允许从单个开放阅读框（sORF）生成表达产物。一个实施方案提供了用于产生一种或多种包含单个开放阅读框插入物的重组蛋白产物的分离或纯化的表达载体; 所述插入片段包含编码信号肽的信号肽核酸序列; 编码第一多肽的第一核酸序列; 编码第一蛋白切割位点的第一插入核酸序列，其中所述第一蛋白切割位点由Pyrococcus的离子蛋白酶基因的内含肽片段或Pyrococcus或Methanococcus的klbA基因或由其衍生的修饰的内含肽片段提供; 和编码第二多肽的第二核酸序列。构建体和方法的某些实施方案采用激烈球菌（Pyrococcus abyssi），激烈火球菌（Pyrococcus furiosus）或火球菌（Pyrococcus horikoshii）OT3的离子蛋白酶基因的内含肽片段; 或激烈球菌（Crrococcus abyssi），激烈火球菌（Pyrococcus furiosus）或季戊糖（Methanococcus jannaschii）的klbA基因的内含肽片段; 或其他内含肽片段。

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