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1. 发明申请

WO2002010414A3 EXPRESSION OF BIOLOGICALLY ACTIVE POLYPEPTIDES IN DUCKWEED 审中-公开
公开(公告)号：WO2002010414A3
公开(公告)日：2002-02-07
申请号：PCT/US2001/023400
申请日：2001-07-26
申请人： BIOLEX, INC. , STOMP, Anne-Marie , DICKEY, Lynn , GASDASKA, John
发明人： STOMP, Anne-Marie , DICKEY, Lynn , GASDASKA, John
IPC分类号： C12N15/82
摘要： Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium.

2. 发明申请

WO2010138631A1 COMPOSITIONS AND METHODS FOR PRODUCTION OF AGLYCOSYLATED PLASMINOGEN 审中-公开
标题翻译：用于生产合成的PLASMINOGEN的组合物和方法
公开(公告)号：WO2010138631A1
公开(公告)日：2010-12-02
申请号：PCT/US2010/036253
申请日：2010-05-26
申请人： BIOLEX THERAPEUTICS, INC. , DICKEY, Lynn, F. , GASDASKA, John, R. , COX, Kevin, M.
发明人： DICKEY, Lynn, F. , GASDASKA, John, R. , COX, Kevin, M.
IPC分类号： C12N9/68 , A61K38/16 , C12N15/82 , C12N5/04 , C12N9/14
CPC分类号： A61K38/168 , C12N9/6435 , C12N15/8257 , C12Y304/21007
摘要： Compositions and methods for producing aglycosylated plasminogen (PLG) polypeptides and fragments and variants thereof are provided. Compositions of the invention include isolated nucleic acid molecules encoding aglycosylated PLG polypeptides in which the asparagine (Asn) residue corresponding to residue Asn-289 of the mature human PLG polypeptide has been substituted with an amino acid residue that does not support N -linked glycosylation at that position of the PLG polypeptide, as well as the aglycosylated PLG polypeptides encoded thereby. Expression constructs comprising these PLG-encoding nucleic acid molecules and transgenic plants comprising these expression constructs are also provided. Methods of the invention comprise introducing a PLG-encoding nucleic acid molecule of the invention into a plant of interest and culturing the plant under conditions to produce the aglycosylated PLG polypeptide. The aglycosylated PLG polypeptide allows for significant increases in production and yield of PLG from a plant-based expression system without comprising the ability of the PLG product to be activated to a polypeptide capable of binding fibrin and having serine protease activity, including biologically active plasmin that is also aglycosylated. The activated aglycosylated plasmin is useful to treat diseases or conditions associated with a thrombus.
摘要翻译：提供了用于产生糖基化纤溶酶原（PLG）多肽及其片段和变体的组合物和方法。本发明的组合物包括分离的编码糖基化PLG多肽的核酸分子，其中对应于成熟人PLG多肽的残基Asn-289的天冬酰胺（Asn）残基已被不支持N-连接糖基化的氨基酸残基取代 PLG多肽的位置，以及由此编码的糖基化的PLG多肽。还提供了包含编码这些编码PLG的核酸分子和包含这些表达构建体的转基因植物的表达构建体。本发明的方法包括将本发明的编码PLG的核酸分子引入感兴趣的植物中，并在产生糖苷基化的PLG多肽的条件下培养该植物。糖基化的PLG多肽允许来自基于植物的表达系统的PLG的产量和产量的显着增加，而不包括PLG产物被活化到能够结合纤维蛋白并具有丝氨酸蛋白酶活性的多肽的能力，包括生物活性纤溶酶，也糖基化。活化的糖基化纤溶酶可用于治疗与血栓相关的疾病或病症。

3. 发明申请

WO2005035768A1 EXPRESSION OF BIOLOGICALLY ACTIVE POLYPEPTIDES IN DUCKWEED 审中-公开
标题翻译：表达生物活性多聚体在DUCKWEED
公开(公告)号：WO2005035768A1
公开(公告)日：2005-04-21
申请号：PCT/US2004/011968
申请日：2004-04-16
申请人： BIOLEX, INC. , DICKEY, Lynn , GASDASKA, John , COX, Kevin , PEELE, Charles, G. , SPENCER, David
发明人： DICKEY, Lynn , GASDASKA, John , COX, Kevin , PEELE, Charles, G. , SPENCER, David
IPC分类号： C12N15/82
CPC分类号： C07K14/56 , C07K16/005 , C07K2317/55 , C12N15/8216 , C12N15/8257
摘要： Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium.
摘要翻译：提供了用于从遗传工程化的浮萍中表达和分泌生物活性多肽的方法，核酸序列和转化的浮萍植物或浮萍结核菌培养物。通过修饰编码多肽的表达盒的核苷酸序列来改善浮萍中重组多肽的表达，从而改善浮萍中的表达。通过将生物活性多肽与指导多肽分泌到培养基中的信号肽连接来提高从浮萍中回收生物活性多肽。

4. 发明申请

WO2005078109A2 EXPRESSION OF PLASMINOGEN AND MICROPLASMINOGEN IN DUCKWEED 审中-公开
标题翻译：表达胰岛素和微管蛋白在DUCKWEED中的表达
公开(公告)号：WO2005078109A2
公开(公告)日：2005-08-25
申请号：PCT/US2005/004245
申请日：2005-02-11
申请人： BIOLEX, INC. , SPENCER, David , DICKEY, Lynn, F. , GASDASKA, John, R. , WANG, Xiaowei , COX, Kevin, M. , PEELE, Charles, G.
发明人： SPENCER, David , DICKEY, Lynn, F. , GASDASKA, John, R. , WANG, Xiaowei , COX, Kevin, M. , PEELE, Charles, G.
IPC分类号： C12N15/82
CPC分类号： C12N15/8257 , C12N9/6435 , C12Y304/21007
摘要： The present invention provides methods and compositions for the production of recombinant plasminogen, microplasminogen, and fragments thereof in a duckweed expression system. It is the novel finding of the present invention that a duckweed expression system may be used to produce high levels of plasminogen and microplasminogen. The duckweed-produced plasminogen and microplasminogen can be activated to produce a polypeptide having protease activity. Thus, the invention encompasses methods for the expression of plasminogen, microplasminogen, and fragments thereof in duckweed, duckweed plants that are transformed with expression cassettes for the expression of plasminogen, microplasminogen, and fragments thereof, and nucleic acids comprising nucleotide sequences encoding plasminogen, microplasminogen, and fragments thereof, where these nucleotide sequences are modified to enhance their expression in duckweed.
摘要翻译：本发明提供了用于在浮萍表达系统中生产重组纤溶酶原，微量血浆原和其片段的方法和组合物。本发明的新发现是，浮萍表达系统可用于产生高水平的纤溶酶原和微量淀粉样蛋白。可以将浮萍产生的纤溶酶原和微量血浆原活化以产生具有蛋白酶活性的多肽。因此，本发明包括在用于表达纤溶酶原，微量血浆原和其片段的表达盒转化的浮萍，浮萍植物中表达纤溶酶原，微量淀粉样蛋白及其片段的方法，以及包含编码纤溶酶原，微量稀释细胞的核苷酸序列的核酸及其片段，其中这些核苷酸序列被修饰以增强其在浮萍中的表达。

5. 发明申请

WO0210414A2 EXPRESSION OF BIOLOGICALLY ACTIVE POLYPEPTIDES IN DUCKWEED 审中-公开
标题翻译：表达生物活性多聚体在DUCKWEED
公开(公告)号：WO0210414A2
公开(公告)日：2002-02-07
申请号：PCT/US0123400
申请日：2001-07-26
申请人： BIOLEX INC , STOMP ANNE MARIE , DICKEY LYNN , GASDASKA JOHN
发明人： STOMP ANNE-MARIE , DICKEY LYNN , GASDASKA JOHN
IPC分类号： C12N15/09 , C07K14/56 , C12N5/10 , C12N9/00 , C12N15/82 , C12P21/02 , C12P21/08 , C12N15/62 , C12N15/67
CPC分类号： C07K14/56 , C12N15/8216 , C12N15/8257
摘要： Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium.
摘要翻译：提供了用于从遗传工程化的浮萍中表达和分泌生物活性多肽的方法，核酸序列和转化的浮萍植物或浮萍结核菌培养物。通过修饰编码多肽的表达盒的核苷酸序列来改善浮萍中重组多肽的表达，从而改善浮萍中的表达。通过将生物活性多肽与指导多肽分泌到培养基中的信号肽连接来提高从浮萍中回收生物活性多肽。

6. 发明申请

WO2005078109A3 EXPRESSION OF PLASMINOGEN AND MICROPLASMINOGEN IN DUCKWEED 审中-公开
标题翻译：杜氏鲍曼不动杆菌的表达及其微丝体表达
公开(公告)号：WO2005078109A3
公开(公告)日：2005-11-10
申请号：PCT/US2005004245
申请日：2005-02-11
申请人： BIOLEX INC , SPENCER DAVID , DICKEY LYNN F , GASDASKA JOHN R , WANG XIAOWEI , COX KEVIN M , PEELE CHARLES G
发明人： SPENCER DAVID , DICKEY LYNN F , GASDASKA JOHN R , WANG XIAOWEI , COX KEVIN M , PEELE CHARLES G
IPC分类号： A01H1/00 , C12N9/68 , C12N15/82
CPC分类号： C12N15/8257 , C12N9/6435 , C12Y304/21007
摘要： The present invention provides methods and compositions for the production of recombinant plasminogen, microplasminogen, and fragments thereof in a duckweed expression system. It is the novel finding of the present invention that a duckweed expression system may be used to produce high levels of plasminogen and microplasminogen. The duckweed-produced plasminogen and microplasminogen can be activated to produce a polypeptide having protease activity. Thus, the invention encompasses methods for the expression of plasminogen, microplasminogen, and fragments thereof in duckweed, duckweed plants that are transformed with expression cassettes for the expression of plasminogen, microplasminogen, and fragments thereof, and nucleic acids comprising nucleotide sequences encoding plasminogen, microplasminogen, and fragments thereof, where these nucleotide sequences are modified to enhance their expression in duckweed.
摘要翻译：本发明提供了用于在浮萍表达系统中生产重组纤溶酶原，微纤溶酶原及其片段的方法和组合物。本发明的新发现是浮萍表达系统可用于产生高水平的纤溶酶原和微纤溶酶原。浮萍产生的纤溶酶原和微纤溶酶原可被活化以产生具有蛋白酶活性的多肽。因此，本发明包括用于在表达纤溶酶原，微纤溶酶原及其片段的表达盒和包含编码纤溶酶原，微纤溶酶原的核苷酸序列的核酸的浮萍，浮萍植物中表达纤溶酶原，微纤溶酶原及其片段的方法及其片段，其中这些核苷酸序列被修饰以增强它们在浮萍中的表达。

7. 发明申请

WO2005035767A1 ALPHA INTERFERON VARIANTS 审中-公开
标题翻译：阿尔法干扰素
公开(公告)号：WO2005035767A1
公开(公告)日：2005-04-21
申请号：PCT/US2004/011965
申请日：2004-04-16
申请人： BIOLEX, INC. , DICKEY, Lynn , GASDASKA, John , COX, Kevin
发明人： DICKEY, Lynn , GASDASKA, John , COX, Kevin
IPC分类号： C12N15/82
CPC分类号： C12N15/8216 , C07K14/56 , C07K2317/55 , C12N15/8257
摘要： The present invention provides biologically active variants of human α-2b- interferon. The variants contain carboxy terminus truncations when compared with the amino acid sequence of full-length human α-2b-interferon. It is the novel finding of the present invention that these truncated variants have the biological activity of full-length human α-2b-interferon. The invention encompasses these biologically active variant α-interferons, as well as polynucleotides encoding these interferons. Expression cassettes comprising these polynucleotides and host cells comprising the expression cassettes are also provided. The invention also provides compositions comprising variant α-interferon polypeptides and a pharmaceutically acceptable carrier.
摘要翻译：本发明提供人α-2b-干扰素的生物活性变体。当与全长人α-2b-干扰素的氨基酸序列相比时，变体含有羧基末端截短。本发明的新发现是这些截短的变体具有全长人α-2b-干扰素的生物学活性。本发明包括这些生物活性变体α-干扰素，以及编码这些干扰素的多核苷酸。还提供了包含这些多核苷酸的表达盒和包含表达盒的宿主细胞。本发明还提供包含变体α-干扰素多肽和药学上可接受的载体的组合物。

8. 发明申请

WO0210414A9 EXPRESSION OF BIOLOGICALLY ACTIVE POLYPEPTIDES IN DUCKWEED 审中-公开
标题翻译：表达生物活性多聚体在DUCKWEED
公开(公告)号：WO0210414A9
公开(公告)日：2003-10-30
申请号：PCT/US0123400
申请日：2001-07-26
申请人： BIOLEX INC , STOMP ANNE-MARIE , DICKEY LYNN , GASDASKA JOHN
发明人： STOMP ANNE-MARIE , DICKEY LYNN , GASDASKA JOHN
IPC分类号： C12N15/09 , C07K14/56 , C12N5/10 , C12N9/00 , C12N15/82 , C12P21/02 , C12P21/08 , C12N15/62 , C12N15/67
CPC分类号： C07K14/56 , C12N15/8216 , C12N15/8257
摘要： Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium.
摘要翻译：提供了用于从遗传工程化的浮萍中表达和分泌生物活性多肽的方法，核酸序列和转化的浮萍植物或浮萍结核菌培养物。通过修饰编码多肽的表达盒的核苷酸序列来改善浮萍中重组多肽的表达，从而改善浮萍中的表达。通过将生物活性多肽与指导多肽分泌到培养基中的信号肽连接来提高从浮萍中回收生物活性多肽。

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