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1. 发明授权

US5024938A Recombinant DNA inserted with hepatitis B virus gene, mammalian cells transformed with cloned viral DNA, and production of hepatitis B virus proteins 失效
标题翻译：用乙型肝炎病毒基因插入的重组DNA，用克隆的病毒DNA转化的哺乳动物细胞，以及产生乙型肝炎病毒蛋白质
公开(公告)号：US5024938A
公开(公告)日：1991-06-18
申请号：US883138
申请日：1986-07-08
申请人： Chikateru Nozaki , Atsushi Miyanohara , Fukusaburo Hamada , Nobuya Ohtomo , Kenichi Matsubara
发明人： Chikateru Nozaki , Atsushi Miyanohara , Fukusaburo Hamada , Nobuya Ohtomo , Kenichi Matsubara
IPC分类号： C12N15/09 , A61K39/29 , C07H21/04 , C07K14/02 , C12N5/10 , C12P21/00 , C12P21/02
CPC分类号： C07K14/005 , C12N2730/10122
摘要： A recombinant DNA comprising 3 fragments of Hepatitis B virus DNA recombined with a vector consisting essentially of 0.31 Kb replication orgin of SV40 DNA inserted into EcoRI cleavage site of Escherichia coli plasmid which is deficient in the 1.426-2.521 Kb region of inhibiting replication in mammalian cells, wherein each HBV DNA fragment is a 3.2 Kb BamHI fragment consisting of 1.9 Kb HBc gene and 1.3 Kb HBs gene, and said HBV DNA fragments are arranged in a head-to-tail tandem relationship wherein the HBc gene positions at the head and the HBs gene positions at the tail, mammalian cells transformed with the recombinant DNA, and a method of production of Hepatitis B virus proteins, i.e. HBsAg and/or HBeAg. These HBV proteins have the same immunological properties as those of the natural HBV proteins originated from human blood plasma and can be used for the preparation of HBV vaccine and diagnostic reagents.
摘要翻译：将包含3个乙型肝炎病毒DNA片段的重组DNA与基本上由插入到在哺乳动物细胞中抑制复制的1.426-2.521Kb区域的缺失的大肠杆菌质粒的EcoRI切割位点的SV40DNA的0.31Kb复制形式的载体重组，其中每个HBV DNA片段是由1.9Kb HBc基因和1.3Kb HBs基因组成的3.2Kb BamHI片段，并且所述HBV DNA片段以头对尾串联关系排列，其中HBc基因位于头部和在尾部的HBs基因位置，用重组DNA转化的哺乳动物细胞，以及生产乙型肝炎病毒蛋白即HBsAg和/或HBeAg的方法。这些HBV蛋白质具有与源自人血浆的天然HBV蛋白质相同的免疫学特性，可用于制备HBV疫苗和诊断试剂。

2. 发明授权

US4778761A Recombinant plasmid inserted with hepatitis B virus gene, yeast transformed with said recombinant plasmid, and production of hepatitis B virus surface antigen 失效
标题翻译：插入乙型肝炎病毒基因的重组质粒，用所述重组质粒转化的酵母，并产生乙型肝炎病毒表面抗原
公开(公告)号：US4778761A
公开(公告)日：1988-10-18
申请号：US4424
申请日：1987-01-20
申请人： Atsushi Miyanohara , Chikateru Nozaki , Fukusaburo Hamada , Akio Toh-e , Nobuya Ohtomo , Kenichi Matsubara
发明人： Atsushi Miyanohara , Chikateru Nozaki , Fukusaburo Hamada , Akio Toh-e , Nobuya Ohtomo , Kenichi Matsubara
IPC分类号： C12N15/09 , A61K39/29 , C07H21/04 , C07K14/02 , C12N15/00 , C12N15/62 , C12N15/80 , C12N15/81 , C12P21/00 , C12P21/02 , C12R1/19 , C12R1/865 , C12N1/00
CPC分类号： C07K14/005 , C12N15/62 , C12N15/80 , C12N15/81 , C07K2319/00 , C07K2319/40 , C12N2730/10122
摘要： A recombinant plasmid inserted with Hepatitis B virus gene, which comprises a plasmid vector containing a yeast gene and an E. coli gene and carrying the expression control region of the repressible acid phosphatase gene of yeast and a Hepatitis B virus gene recombined thereto under control of the phosphatase promoter, a transformed yeast which is prepared by transforming a yeast with the recombinant plasmid, and a method of the production of Hepatitis B virus surface antigen in a large scale by culturing the transformed yeast in a medium. The Hepatitis B virus surface antigen prepared by the present invention has the same immunological properties as those of the natural antigen from human blood plasma and is useful for the preparation of Hepatitis B virus vaccine and diagnostic reagents.
摘要翻译：插入乙型肝炎病毒基因的重组质粒，其包含含有酵母基因和大肠杆菌基因的质粒载体，并携带酵母的可压制酸性磷酸酶基因的表达控制区和重组的乙型肝炎病毒基因的控制区磷酸酶启动子，通过用重组质粒转化酵母制备的转化酵母以及通过在培养基中培养转化的酵母大规模生产乙型肝炎病毒表面抗原的方法。本发明制备的乙型肝炎病毒表面抗原具有与人血浆天然抗原相同的免疫学特性，可用于制备乙型肝炎病毒疫苗和诊断试剂。

3. 发明授权

US4162192A Method for purification of HBs antigen 失效
标题翻译： HBs抗原纯化方法
公开(公告)号：US4162192A
公开(公告)日：1979-07-24
申请号：US946733
申请日：1978-09-28
申请人： Kyosuke Mizuno , Atsushi Miyanohara , Yoshimitsu Ishihara , Nobuya Ohtomo
发明人： Kyosuke Mizuno , Atsushi Miyanohara , Yoshimitsu Ishihara , Nobuya Ohtomo
IPC分类号： A61K39/00 , C07K14/02 , A61K39/12
CPC分类号： C07K14/005 , A61K39/00 , C12N2730/10122
摘要： An improved method for purification of HBs antigen comprising passing a partially purified HBs antigen prepared from blood plasma or serum through a column filled with an anion exchanger which may be equilibrated with a buffer solution having a specific ionic strength and pH level, and further passing the resulting effluent through a column filled with a cation exchanger which may be equilibrated with a buffer solution having a specific ionic strength and pH level. The HBs antigen obtained by the present invention is highly purified and is useful for preparing vaccine for preventing Viral Hepatitis Type B.
摘要翻译：一种用于纯化HBs抗原的改进方法，包括将由血浆或血清制备的部分纯化的HBs抗原通过填充有阴离子交换剂的柱，其可以用具有特定离子强度和pH水平的缓冲溶液平衡，并进一步通过得到的流出物通过填充有阳离子交换剂的柱，其可以用具有特定离子强度和pH水平的缓冲溶液平衡。本发明获得的HBs抗原是高度纯化的，可用于制备用于预防乙型肝炎病毒的疫苗。

4. 发明授权

US5707862A Shuttle vector 失效
标题翻译：班车矢量
公开(公告)号：US5707862A
公开(公告)日：1998-01-13
申请号：US622344
申请日：1990-12-05
申请人： Atsushi Miyanohara , Akio Toh-e , Kenichi Matsubara
发明人： Atsushi Miyanohara , Akio Toh-e , Kenichi Matsubara
IPC分类号： C12N15/09 , A61K39/29 , C07H21/04 , C07K14/02 , C12N1/19 , C12N15/00 , C12N15/62 , C12N15/81 , C12P19/34 , C12P21/00 , C12R1/19 , C12R1/865
CPC分类号： C07K14/005 , C12N15/62 , C12N15/81 , C07K2319/00 , C12N2730/10122
摘要： A shuttle vector containing an yeast gene and an E. coli gene and carrying the expression control region of acid phosphatase gene of yeast, which can be recombined with various genes under control of the phosphatase promoter, to give various recombinant plasmids. The shuttle vector is useful in genetic engineering industries.
摘要翻译：含有酵母基因和大肠杆菌基因的穿梭载体，并携带可与磷酸酶启动子控制下的各种基因重组的酵母酸性磷酸酶基因的表达控制区，得到各种重组质粒。穿梭载体在基因工程行业很有用。

5. 发明授权

US06451304B1 Method for retrovirus vector production by separated gag and pol expression 失效
标题翻译：通过分离的gag和pol表达产生逆转录病毒载体的方法
公开(公告)号：US06451304B1
公开(公告)日：2002-09-17
申请号：US09265013
申请日：1999-03-09
申请人： Theodore Friedmann , Atsushi Miyanohara
发明人： Theodore Friedmann , Atsushi Miyanohara
IPC分类号： A01N6300
CPC分类号： C12N15/86 , C12N2740/13043 , C12N2740/16043
摘要： The invention provides a system for production of retroviruses which are replication incompetent. In the system, gag and pol retroviral structural proteins are expressed separately by different plasmids in a packaging cell line. Separate gag and pol expressing plasmids are provided, as are packaging cell lines containing such plasmids. Retrovirus products of the retroviral vector production system, including chimeric retroviruses, are also provided.
摘要翻译：本发明提供了一种用于生产复制无能力的逆转录病毒的系统。在系统中，gag和pol逆转录病毒结构蛋白在包装细胞系中由不同质粒分开表达。提供单独的gag和pol表达质粒，以及含有这种质粒的包装细胞系也是如此。还提供了逆转录病毒载体生产系统的逆转录病毒产品，包括嵌合逆转录病毒。

6. 发明授权

US06171862B2 Transfection in serum-containing media 失效
标题翻译：含血清培养基中的转染
公开(公告)号：US06171862B2
公开(公告)日：2001-01-09
申请号：US09052797
申请日：1998-03-31
申请人： Akihiro Abe , Atsushi Miyanohara , Theodore Friedmann
发明人： Akihiro Abe , Atsushi Miyanohara , Theodore Friedmann
IPC分类号： C12N1587
CPC分类号： C12N15/87 , A61K48/00 , C12N15/88
摘要： Transfection of host cells cultured in the presence of serum is increased by adding VSV-G or polybrene to a nucleic acid-lipid complex or culture medium prior to transfection.
摘要翻译：在转染前，通过向核酸 - 脂质复合物或培养基中加入VSV-G或聚凝胺来增加在血清存在下培养的宿主细胞的转染。

7. 发明授权

US5739018A Packaging cell lines for pseudotyped retroviral vectors 失效
标题翻译：包装用于假型逆转录病毒载体的细胞系
公开(公告)号：US5739018A
公开(公告)日：1998-04-14
申请号：US694652
申请日：1996-08-07
申请人： Atsushi Miyanohara , Jiing-Kuan Yee , Shin-Tai Chen , Charles Edward Prussak , Theodore Friedmann
发明人： Atsushi Miyanohara , Jiing-Kuan Yee , Shin-Tai Chen , Charles Edward Prussak , Theodore Friedmann
IPC分类号： C12N15/63 , C12N15/867 , C12N5/10 , C12N15/86
CPC分类号： C12N15/86 , C12N15/635 , C12N2740/13052 , C12N2830/002 , C12N2830/006 , C12N2830/15 , C12N2830/30 , C12N2840/20 , C12N2840/203
摘要： The present invention features packaging cell lines and recombinant retroviral particles produced thereby, particularly pseudotyped retroviral particles. Preferably, the packaging cell lines are derived from HeLa, Cf2Th, D17, MDCK, or BHK cells, most preferably from Cf2Th cells. Retroviral particles are produced by inducibly expressing an envelope protein of interest (e.g., a retroviral envelope or the envelope protein of vesicular stomatitis virus (VSV G)). Inducible expression of the envelope protein is accomplished by operably linking an envelope protein-encoding nucleotide sequence to an inducible promoter (e.g., a promoter composed of a minimal promoter linked to multiple copies of tetO, the binding site for the tetracycline repressor (tetR) of the Escherichia coli, tetracycline resistance operon Tn10). Expression from the inducible promoter is regulated by a multi-chimeric transactivating factor, composed of a first ligand-binding domain that negatively regulates transcription from the inducible promoter (e.g., a prokaryotic tetracycline repressor polypeptide (tetR)), a transcriptional activation domain, and a second ligand-binding domain (e.g., a ligand-binding domain of a steroid receptor, preferably an estrogen receptor (ER)).
摘要翻译：本发明的特征在于包装细胞系和由此产生的重组逆转录病毒颗粒，特别是假型逆转录病毒颗粒。优选地，包装细胞系衍生自HeLa，Cf2Th，D17，MDCK或BHK细胞，最优选来自Cf2Th细胞。逆转录病毒颗粒通过诱导表达感兴趣的包膜蛋白（例如，逆转录病毒包膜或水泡性口炎病毒的包膜蛋白（VSV G））产生。通过将包膜蛋白编码核苷酸序列可操作地连接到诱导型启动子（例如，由与tetO的多拷贝连接的最小启动子，四环素阻遏物（tetR）的结合位点）组成的启动子来实现包膜蛋白的诱导表达大肠杆菌，四环素抗性操纵子Tn10）。来自诱导型启动子的表达由多嵌合反式激活因子调节，所述多嵌合反式激活因子由负调节来自诱导型启动子的转录的第一配体结合结构域（例如，原核四环素阻遏物多肽（tetR）），转录激活结构域和第二配体结合结构域（例如，类固醇受体的配体结合结构域，优选雌激素受体（ER））。

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