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公开(公告)号：US06174724B1
公开(公告)日：2001-01-16
申请号：US08435951
申请日：1995-05-04
IPC分类号： C12N1582
CPC分类号： C12N15/8205 , C12N15/8216
摘要： This invention relates to chimeric genes which are capable of being expressed in plant cells. Such genes contain (a) a promoter region derived a gene which is expressed in plant cells, such as the nopaline synthase gene; (b) a coding or structural sequence which is heterologous with respect to the promoter region; and (c) an appropriate 3′ non-translated region. Such genes have been used to create antibiotic-resistant plant cells; they are also useful for creating herbicide-resistant plants, and plants which contain mammalian polypeptides.
摘要翻译： 本发明涉及能够在植物细胞中表达的嵌合基因。 这些基因含有（a）衍生自在植物细胞中表达的基因的启动子区，例如胭脂碱合酶基因; （b）相对于启动子区域是异源的编码或结构序列; 和（c）适当的3'非翻译区域。 已经使用这样的基因来产生抗生素抗性植物细胞; 它们也可用于产生除草剂抗性植物和含有哺乳动物多肽的植物。
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公开(公告)号：US08334139B1
公开(公告)日：2012-12-18
申请号：US06783336
申请日：1985-10-04
CPC分类号： C12N15/8205
摘要： This invention relates to several plasmids which are useful for genetically transforming plant cells. A first plasmid, such as pMON120, contains a T-DNA border, one or more marker genes, a unique cleavage site, and a region of Ti plasmid homology. A gene which is expressed in plant cells may be inserted into this plasmid to obtain a derivative plasmid, such as pMON128 which expresses neomycin phosphotransferase in plant cells. The derivative plasmid is inserted into a suitable microorganism, such as A. tumefaciens which contains a Ti plasmid. The inserted plasmids recombine with Ti plasmids to form co-integrate plasmids. Only a single crossover event is required to create the desired co-integrate plasmid. A. tumefaciens cells with co-integrate plasmids are selected and co-cultured with plant cells. The co-integrate Ti plasmids enter the plant cells and insert a segment of T-DNA which does not contain tumorigenic genes into the plant genome. The transformed plant cell(s) may be regenerated into a morphologically normal plant which will pass the inserted gene(s) to its descendants.
摘要翻译： 本发明涉及可用于遗传转化植物细胞的几种质粒。 第一种质粒如pMON120含有T-DNA边界，一个或多个标记基因，独特的切割位点和Ti质粒同源性的区域。 可以将在植物细胞中表达的基因插入该质粒中，得到在植物细胞中表达新霉素磷酸转移酶的pMON128等衍生质粒。 将衍生质粒插入合适的微生物，如含有Ti质粒的根瘤土壤杆菌。 插入的质粒与Ti质粒重组以形成共同整合质粒。 只需要一个交叉事件来创建所需的共同整合质粒。 选择具有共同整合质粒的根癌农杆菌细胞并与植物细胞共培养。 共同整合的Ti质粒进入植物细胞，并将不含致瘤基因的一段T-DNA插入植物基因组。 转化的植物细胞可以再生成形态上正常的植物，其将插入的基因传递给其后代。
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公开(公告)号：US4948731A
公开(公告)日：1990-08-14
申请号：US893482
申请日：1986-08-05
申请人： Lee Gehrke , Robert T. Fraley , Stephen G. Rogers
发明人： Lee Gehrke , Robert T. Fraley , Stephen G. Rogers
摘要： A recombinant plasmid which may be used for propagation of cloned cDNAs and also for the in vitro synthesis of RNA which is an exact copy of the natural sequence, wherein the transcript is devoid of vector-derived sequence. The novel vector was generated de novo by genetic engineering procedures using a synthetic double strand oligodeoxyribonucleotide fragment and a larger DNA fragment derived from plasmid pSP64. The vector, plasmid pHST-O, carried by Escherichia coli HB101, deposited with the ATCC on Dec. 30, 1985 and designated ATCC 53381, is distinguished from other vectors containing the SP6 promoter by the following characteristics: it contains a unique site for the restriction endonuclease BglII; the engineered GBlII site overlaps the downstream border of the SP6 promoter sequence; and the presence and positioning of the BglII restriction site permit insertion of cDNA molecules in such a way that transcription by the SP6 RNA polymerase begins exactly at the 5' terminus of the RNA, providing that the 5' terminal nucleotide of the mRNA transcript is a guanosine (G) residue, thus excluding the transcription of nucleotides derived from the vector. The novel vector permits synthesis of RNA molecules which have a defined 5' terminus and which are devoid of vector-derived sequence. The vector has potential use in research in molecular biology and in the in vitro production of RNA and proteins.
摘要翻译： 可用于繁殖克隆cDNA的重组质粒以及用于体外合成RNA的重组质粒，其是天然序列的精确拷贝，其中转录本缺乏载体衍生的序列。 通过使用合成双链寡脱氧核糖核苷酸片段和源自质粒pSP64的较大DNA片段的遗传工程程序从头生成新载体。 1985年12月30日保藏于ATCC并命名为ATCC 53381的大肠杆菌HB101携带的载体质粒pHST-O与含有SP6启动子的其他载体具有以下特征的区别：其含有 限制性内切核酸酶BglII; 设计的GBlII位点与SP6启动子序列的下游边界重叠; 并且BglII限制性位点的存在和定位允许插入cDNA分子，使得由SP6 RNA聚合酶转录正好位于RNA的5'末端，条件是mRNA转录物的5'末端核苷酸是 鸟苷（G）残基，因此不包括衍生自载体的核苷酸的转录。 该新载体允许合成具有定义的5'末端并且没有载体衍生序列的RNA分子。 该载体在分子生物学和RNA和蛋白质的体外生产研究中具有潜在的用途。
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公开(公告)号：US06608241B1
公开(公告)日：2003-08-19
申请号：US06917027
申请日：1986-10-09
IPC分类号： C12N1533
CPC分类号： C12N15/1131 , A01H1/00 , C07K14/005 , C12N15/8283 , C12N15/87 , C12N2750/12022 , C12N2770/14022 , C12N2770/40022
摘要： The invention involves recombinant, double-stranded DNA that contains a promoter which functions in plant cells to cause the production of RNA sequences of a plant virus, a DNA sequence that causes the production of an RNA sequence encoding the coat protein of said plant virus, and a 3′ non-translated region which functions in plant cells to cause the addition of polyadenylated nucleotides to the 3′ end of said RNA sequence; which double-stranded DNA can be used in a method for genetically transforming plants to produce genetically transformed plant cells and plants that are resistant to virus infection.
摘要翻译： 本发明涉及包含在植物细胞中起作用以产生植物病毒RNA序列的启动子的重组双链DNA，导致产生编码所述植物病毒外壳蛋白的RNA序列的DNA序列， 和3'非翻译区，其在植物细胞中起作用以引起多聚腺苷酸核苷酸加入所述RNA序列的3'末端; 该双链DNA可用于遗传转化植物以产生对病毒感染有抗性的遗传转化的植物细胞和植物的方法。
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公开(公告)号：US5530196A
公开(公告)日：1996-06-25
申请号：US300029
申请日：1994-09-02
IPC分类号： C07K14/61 , C12N15/34 , C12N15/52 , C12N15/82 , C12N15/83 , C12N15/84 , A01H5/00 , A01H5/10 , C12N5/04
CPC分类号： C07K14/61 , C12N15/52 , C12N15/8216
摘要： In one aspect the present invention relates to the use of viral promoters in the expression of chimeric genes in plant cells. In another aspect this invention relates to chimeric genes which are capable of being expressed in plant cells, which utilize promoter regions derived from viruses which are capable of infecting plant cells. One such virus comprises the cauliflower mosaic virux (CaMV). Two different promoter regions have been derived from the CaMV genome and ligated to heterologous coding sequences to form chimeric genes. These chimeric genes have been shown to be expressed in plant cells. This invention also relates to plant cells, plant tissue, and differentiated plants which contain and express the chimeric genes of this invention.
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公开(公告)号：US5034322A
公开(公告)日：1991-07-23
申请号：US333802
申请日：1989-04-05
CPC分类号： C07K14/61 , C12N15/52 , C12N15/8216
摘要： This invention relates to chimeric genes which are capable of being expressed in plant cells. Such genes contain (a) a promoter region derived in a gene which is expressed in plant cells, such as the nopaline synthase gene; (b) a coding or structural sequence which is heterologous with respect to the promoter region; and (c) an appropriate 3' non-translated region. Such genes have been used to create antibiotic-resistant plant cells; they are also useful for creating herbicide-resistant plants, and plants which contain mammalian polypeptides.
摘要翻译： 本发明涉及能够在植物细胞中表达的嵌合基因。 这些基因含有（a）在植物细胞中表达的基因中衍生的启动子区，例如胭脂碱合酶基因; （b）相对于启动子区域是异源的编码或结构序列; 和（c）适当的3'非翻译区域。 已经使用这样的基因来产生抗生素抗性植物细胞; 它们也可用于产生除草剂抗性植物和含有哺乳动物多肽的植物。
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公开(公告)号：US4940835A
公开(公告)日：1990-07-10
申请号：US879814
申请日：1986-07-07
IPC分类号： C12N15/82
CPC分类号： C12N15/8275
摘要： This invention involves a cloning or expression vector comprising a gene which encodes 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) polypeptide which, when expressed in a plant cell contains a chloroplast transit peptide which allows the polypeptide, or an enzymatically active portion thereof, to be transported from the cytoplasm of the plant cell into a chloroplast in the plant cell, and confers a substantial degree of glyphosate resistance upon the plant cell and plants regenerated therefrom.The EPSPS coding sequence may be ligated to a strong promoter, such as the 35S promoter from cauliflower mosaic virus, to create a chimeric gene. Such genes can be inserted into plant transformation vectors, and subsequently introduced into plant cells. Plant cells transformed using such genes and plants regenerated therefrom have been shown to exhibit a substantial degree of glyphosate resistance.
摘要翻译： 本发明涉及包含编码5-烯醇丙酮酸莽草酸-3-磷酸合成酶（EPSPS）多肽的基因的克隆或表达载体，其在植物细胞中表达时含有允许多肽的叶绿体转运肽或其酶活性部分， 从植物细胞的细胞质转移到植物细胞中的叶绿体中，并且赋予植物细胞和植物再生的植物大量的草甘膦抗性。 EPSPS编码序列可以连接到强启动子，例如来自花椰菜花叶病毒的35S启动子，以产生嵌合基因。 可将这些基因插入植物转化载体中，随后导入植物细胞。 已经显示使用从其再生的这些基因和植物转化的植物细胞表现出相当程度的草甘膦抗性。
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公开(公告)号：US08273954B1
公开(公告)日：2012-09-25
申请号：US06793486
申请日：1985-10-30
CPC分类号： C12N15/8205
摘要： This invention relates to genetically transformed, non-tumorous plant cells. A modified Ti plasmid is created which contains a left T-DNA border, one or more desired genes, and a right T-DNA border. This region does not contain tumorigenic or phytohormone-altering genes. The Ti plasmid is inserted into plant cells, where the T-DNA region is transferred into the plant genome. The transformed plant cells may be regenerated into morphologically normal plants which will pass the desired gene(s) to their descendants.
摘要翻译： 本发明涉及遗传转化的非肿瘤植物细胞。 产生修饰的Ti质粒，其包含左T-DNA边界，一个或多个所需基因和右T-DNA边界。 该区域不含有致瘤性或植物激素改变基因。 将Ti质粒插入到植物细胞中，其中T-DNA区被转移到植物基因组中。 转化的植物细胞可以再生成形态上正常的植物，其将通过所需基因到其后代。
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公开(公告)号：US6147278A
公开(公告)日：2000-11-14
申请号：US261770
申请日：1999-03-03
申请人： Stephen G. Rogers , Leslie Brand , Robert B. Horsch , Robert T. Fraley , James Scott Elmer , David Bisaro
发明人： Stephen G. Rogers , Leslie Brand , Robert B. Horsch , Robert T. Fraley , James Scott Elmer , David Bisaro
CPC分类号： C12N15/8203
摘要： The invention relates to novel plant plasmid vectors comprising geminivirus DNA or a portion thereof having inserted therein a heterologous DNA sequence or gene, to processes and DNA intermediates useful in producing said vectors and to methods utilizing such vectors to replicate and express heterologous DNA sequences or genes in plants. In some embodiments, methods and compositions are provided for Ti plasmid delivery of these novel vectors into plants. In other embodiments, methods and compositions are provided which allow for the generation of geminivirus DNA containing plant plasmids in stably transformed plants. In still other embodiments, methods and compositions are provided for replicating and expressing heterologous DNA sequences or genes in plants employing the geminivirus DNA containing vectors of the present invention without causing disease symptoms.
摘要翻译： 本发明涉及包含双子病毒DNA或其中插入有异源DNA序列或基因的部分的新型植物质粒载体，可用于生产所述载体的方法和DNA中间体，以及利用此类载体复制和表达异源DNA序列或基因的方法 在植物中。 在一些实施方案中，提供了将这些新载体的Ti质粒递送到植物中的方法和组合物。 在其它实施方案中，提供了允许在稳定转化的植物中产生含有植物质粒的双系病毒DNA的方法和组合物。 在另外的其它实施方案中，提供了方法和组合物，用于在不引起疾病症状的情况下使用含有本发明的含有双重病毒DNA的载体在植物中复制和表达异源DNA序列或基因。
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公开(公告)号：US5352605A
公开(公告)日：1994-10-04
申请号：US146621
申请日：1993-10-28
IPC分类号： C07K14/61 , C12N15/34 , C12N15/52 , C12N15/82 , C12N15/83 , C12N15/84 , C12N5/00 , C07H21/04 , C12N15/00
CPC分类号： C07K14/61 , C12N15/52 , C12N15/8216
摘要： In one aspect the present invention relates to the use of viral promoters in the expression of chimeric genes in plant cells. In another aspect this invention relates to chimeric genes which are capable of being expressed in plant cells, which utilize promoter regions derived from viruses which are capable of infecting plant cells. One such virus comprises the cauliflower mosaic virus (CaMV). Two different promoter regions have been derived from the CaMV genome and ligated to heterologous coding sequences to form chimeric genes. These chimeric genes have been shown to be expressed in plant cells. This invention also relates to plant cells, plant tissue, and differentiated plants which contain and express the chimeric genes of this invention.
摘要翻译： 一方面，本发明涉及病毒启动子在植物细胞中嵌合基因表达中的应用。 在另一方面，本发明涉及能够在植物细胞中表达的嵌合基因，其利用能够感染植物细胞的来源于病毒的启动子区域。 一种这样的病毒包括花椰菜花叶病毒（CaMV）。 已经从CaMV基因组衍生出两个不同的启动子区，并连接到异源编码序列以形成嵌合基因。 已经显示这些嵌合基因在植物细胞中表达。 本发明还涉及含有和表达本发明的嵌合基因的植物细胞，植物组织和分化植物。
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