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    • 1. 发明授权
    • Nucleic acid molecules encoding endonucleases and methods of use thereof
    • 编码核酸内切酶的核酸分子及其使用方法
    • US07273739B2
    • 2007-09-25
    • US11417448
    • 2006-05-03
    • Hal S. PadgettAndrew A. Vaewhongs
    • Hal S. PadgettAndrew A. Vaewhongs
    • C12N9/16C07K14/00C12P21/06C12Q1/44C12N15/00C12N5/10C12N1/21C07H21/04
    • C12N15/102C07K14/005C12N9/22C12N15/1027C12N15/8203C12N15/8257C12N2770/00022
    • We describe here an in vitro method of increasing complementarity in a heteroduplex polynucleotide sequence. The method uses annealing of opposite strands to form a polynucleotide duplex with mismatches. The heteroduplex polynucleotide is combined with an effective amount of enzymes having strand cleavage activity, 3′ to 5′ exonuclease activity, and polymerase activity, and allowing sufficient time for the percentage of complementarity to be increased within the heteroduplex. Not all heteroduplex polynucleotides will necessarily have all mismatches resolved to complementarity. The resulting polynucleotide is optionally ligated. Several variant polynucleotides result. At sites where either of the opposite strands has templated recoding in the other strand, the resulting percent complementarity of the heteroduplex polynucleotide sequence is increased. The parent polynucleotides need not be cleaved into fragments prior to annealing heterologous strands. Therefore, no reassembly is required.
    • 我们在这里描述了增加异源双链多核苷酸序列中的互补性的体外方法。 该方法使用相反链的退火以形成具有错配的多核苷酸双链体。 将异源双链多核苷酸与有效量的具有链切割活性,3'至5'核酸外切酶活性和聚合酶活性的酶组合,并且允许足够的时间在异源双链体内增加互补百分比。 并非所有的异源双链多核苷酸都必然具有解决互补性的所有错配。 任选地连接得到的多核苷酸。 产生几种变异多核苷酸。 在相对链中的任一个在另一条链中模板记录的位点,异源双链多核苷酸序列的所得百分比互补性增加。 在退火异源链之前,母体多核苷酸不需要切割成片段。 因此,不需要重新组装。
    • 6. 发明申请
    • NUCLEIC ACID MOLECULES ENCODING MISMATCH ENDONUCLEASES AND METHODS OF USE THEREOF
    • 编码缺失缺失突变体的核酸分子及其使用方法
    • US20080145913A1
    • 2008-06-19
    • US11860465
    • 2007-09-24
    • Hal S. PadgettAndrew A. Vaewhongs
    • Hal S. PadgettAndrew A. Vaewhongs
    • C12N9/18
    • C12N15/102C07K14/005C12N9/22C12N15/1027C12N15/8203C12N15/8257C12N2770/00022
    • We describe here an in vitro method of increasing complementarity in a heteroduplex polynucleotide sequence. The method uses annealing of opposite strands to form a polynucleotide duplex with mismatches. The heteroduplex polynucleotide is combined with an effective amount of enzymes having strand cleavage activity, 3′ to 5′ exonuclease activity, and polymerase activity, and allowing sufficient time for the percentage of complementarity to be increased within the heteroduplex. Not all heteroduplex polynucleotides will necessarily have all mismatches resolved to complementarity. The resulting polynucleotide is optionally ligated. Several variant polynucleotides result. At sites where either of the opposite strands has templated recoding in the other strand, the resulting percent complementarity of the heteroduplex polynucleotide sequence is increased. Also described are mismatch endonucleases suitable for use in the process.
    • 我们在这里描述了增加异源双链多核苷酸序列中的互补性的体外方法。 该方法使用相反链的退火以形成具有错配的多核苷酸双链体。 将异源双链多核苷酸与有效量的具有链切割活性,3'至5'核酸外切酶活性和聚合酶活性的酶组合,并且允许足够的时间在异源双链体内增加互补百分比。 并非所有的异源双链多核苷酸都必然具有解决互补性的所有错配。 任选地连接得到的多核苷酸。 产生几种变异多核苷酸。 在相对链中的任一个在另一条链中模板记录的位点,异源双链多核苷酸序列的所得百分比互补性增加。 还描述了适用于该方法的不匹配核酸内切酶。
    • 8. 发明授权
    • Cytoplasmic gene inhibition or gene expression in transfected plants by a tobraviral vector
    • 通过载体对转染植物进行细胞质基因抑制或基因表达
    • US07498480B2
    • 2009-03-03
    • US10634221
    • 2003-08-04
    • Peter D. RobertsMonto H. KumagaiAndrew A. Vaewhongs
    • Peter D. RobertsMonto H. KumagaiAndrew A. Vaewhongs
    • C12N15/83C12N5/04C12N15/82
    • A01H1/04A01H5/12C12N15/1034C12N15/8261C12Q1/68Y02A40/146
    • This invention is directed to a monopartite RNA viral vector comprising modified tobravirus RNA-1 comprising an inserted foreign RNA sequence. This invention is also directed to a bipartite RNA viral vector derived from a tobravirus, wherein the vector comprises one or more foreign RNA sequences. The invention is directed to a method of silencing one or more endogenous plant host genes and a method of simultaneously silencing a plant host gene and expressing a foreign gene in a plant host. Such methods comprise infecting a plant host with a bipartite vector comprising modified tobravirus RNA-1 and RNA-2. The invention is further directed to a method of compiling a plant functional gene profile, a method of changing the phenotype or biochemistry of a plant host, and a method of determining the presence of a trait in a plant host, using a monopartite or bipartite viral vector derived from a tobravirus.
    • 本发明涉及包含经插入的外源RNA序列的修饰的载体RNA-1的单分子RNA病毒载体。 本发明还涉及衍生自托巴威病毒的二分之一RNA病毒载体,其中所述载体包含一个或多个外源RNA序列。 本发明涉及沉默一种或多种内源性植物宿主基因的方法和同时使植物宿主基因沉默并在植物宿主中表达外源基因的方法。 这样的方法包括用包含修饰的tobravirus RNA-1和RNA-2的二重载体感染植物宿主。 本发明进一步涉及一种编码植物功能基因谱的方法,改变植物宿主的表型或生物化学的方法,以及使用单一或二分之一病毒测定植物宿主中性状的存在的方法 衍生自Tobravirus的载体。