公开(公告)号：US5091309A

公开(公告)日：1992-02-25

申请号：US395007

申请日：1989-08-17

申请人： Sondra Schlesinger ,  Henry V. Huang ,  Robin Levis ,  Barbara Weiss ,  Manuel Tsiang

发明人： Sondra Schlesinger ,  Henry V. Huang ,  Robin Levis ,  Barbara Weiss ,  Manuel Tsiang

IPC分类号： C07K14/18 ,  C12N15/86

CPC分类号： C07K14/005 ,  C12N15/86 ,  C12N2770/36122 ,  C12N2770/36143 ,  Y10S435/948

摘要： The present invention relates to RNA vectors comprising Sindbis virus DI RNA having inserted therein heterologous RNA. The vectors of the present invention provide for the cloning and expression of heterologous RNA in eucaryotic cells as well as the packaging of heterologous RNA into viral particles in the presence of helper virus. Transformed cells and genetically altered alphavirus particles comprising alphavirus DI RNA having inserted therein heterologous RNA are also provided.

摘要翻译： 本发明涉及包含插入有异源RNA的辛德比斯病毒DI RNA的RNA载体。 本发明的载体提供了在真核细胞中克隆和表达异源RNA以及在辅助病毒存在下将异源RNA包装到病毒颗粒中。 还提供了转化的细胞和遗传改变的甲病毒颗粒，其包含插入其中的异源RNA的甲病毒属RNA。

公开(公告)号：US5851757A

公开(公告)日：1998-12-22

申请号：US774406

申请日：1996-12-31

申请人： Paul D. Olivo ,  Sondra Schlesinger

发明人： Paul D. Olivo ,  Sondra Schlesinger

IPC分类号： C07K14/08 ,  C12N5/10 ,  C12N15/09 ,  C12Q1/34 ,  C12Q1/66 ,  C12Q1/68 ,  C12Q1/70 ,  G01N33/569 ,  C12Q1/02 ,  C12Q1/04

CPC分类号： C07K14/005 ,  C12Q1/6897 ,  G01N33/56983 ,  C12N2760/18822 ,  G01N2333/085 ,  G01N2333/18 ,  Y10S435/975

摘要： Cell lines and methods are disclosed for detecting the presence of RNA viruses in a specimen. The cell lines are stably transformed with a DNA molecule that includes a promoter capable of being recognized by the DNA dependent RNA polymerase of the cell capable of directing the transcription of a cDNA of a structurally defective RNA virus genome operably coupled to the promoter. The cDNA contains a structural coding sequence encoding a selected reporter gene product. The RNA molecules transcribed by the DNA dependent RNA polymerase are not capable of causing the translation of the reporter gene in the cell except when an active related virus that provides the necessary trans-acting enzymes to cause the increased replication of the RNA containing the reporter gene which is then translated into the reporter gene product is provided. Methods utilizing the cell lines of this invention to detect RNA viruses in a specimen by incubating the specimen with the cell line and assaying for expression of the reporter gene and a kit containing a supply of the cells and a supply of the reagents necessary for the detection of the reporter gene product are also provided.

摘要翻译： 公开了用于检测样品中RNA病毒的存在的细胞系和方法。 细胞系被包括能够被能够引导结构有缺陷的RNA病毒基因的cDNA的转录的细胞的DNA依赖性RNA聚合酶识别的启动子的DNA分子稳定转化，所述基因可操作地连接到启动子上。 cDNA含有编码所选报道基因产物的结构编码序列。 由DNA依赖性RNA聚合酶转录的RNA分子不能引起报告基因在细胞中的翻译，除了当提供必需的反式作用酶以引起含有报告基因的RNA的复制增加的活性相关病毒时 然后将其翻译成报告基因产物。 利用本发明的细胞系的方法通过将样品与细胞系一起孵育并测定报告基因的表达和含有细胞供应的试剂盒和检测所必需的试剂的供应来检测标本中的RNA病毒 的报告基因产物也被提供。

公开(公告)号：US5217879A

公开(公告)日：1993-06-08

申请号：US815602

申请日：1991-12-27

申请人： Henry V. Huang ,  Robin Levis ,  Charles M. Rice ,  Sondra Schlesinger ,  Ping Shen ,  Cheng Xiong

发明人： Henry V. Huang ,  Robin Levis ,  Charles M. Rice ,  Sondra Schlesinger ,  Ping Shen ,  Cheng Xiong

IPC分类号： C12N7/01 ,  C12N15/86

CPC分类号： C12N15/86 ,  C12N2760/18843

摘要： Compositions and methods are disclosed for expressing heterologous coding sequences in host cells. The compositions include infectious Sindbis virus RNA molecules containing at least one heterologous coding sequence inserted within the structural region of the Sindbis virus genome. RNA molecules consisting essentially of a Sindbis virus junction region are also provided. Methods utilizing the novel compositions of the present invention to express heterologous coding sequences in transformed host cells are provided. Methods for producing infectious Sindbis virus particles containing infectious Sindbis virus RNA molecules comprising at least one heterologous coding sequence are provided.This invention was made with government support under grant numbers AI24134, AI11377 and AG05681 awarded by the National Institutes of Health. The government has certain rights in the invention.

摘要翻译： 公开了用于在宿主细胞中表达异源编码序列的组合物和方法。 组合物包括感染性的辛德比斯病毒RNA分子，其含有插入在辛德比斯病毒基因组的结构区域内的至少一个异源编码序列。 还提供了基本上由辛德毕斯病毒连接区组成的RNA分子。 提供了利用本发明新组合物在转化的宿主细胞中表达异源编码序列的方法。 提供了包含至少一种异源编码序列的含感染性辛德毕斯病毒RNA分子的感染性辛德比斯病毒颗粒的方法。