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    • 4. 发明授权
    • Engineering plant thioesterases for altered substrate specificity
    • 工程植物硫酯酶改变底物特异性
    • US5955329A
    • 1999-09-21
    • US440845
    • 1995-05-15
    • Ling YuanKatayoon DeheshJean KridlVic Knauf
    • Ling YuanKatayoon DeheshJean KridlVic Knauf
    • C12N9/16C12N15/82C12N15/00C07H21/04
    • C12N15/8247C12N9/16
    • Methods of altering substrate specificity of plant acyl-ACP thioesterases, and engineered plant acyl-ACP thioesterases so produced are provided. The C-terminal two thirds portion of plant thioesterases is identified as desirable for such modifications. DNA sequences and constructs for expression of engineered thioesterases, as well as the novel thioesterases produced therefrom are also provided. Such DNA sequences may be used for expression of the engineered thioesterases in host cells, particularly seed cells of oilseed crop plants, for the modification of fatty acid composition. A C12 preferring plant acyl-ACP thioesterase described herein may be altered to obtain a plant thioesterase having approximately equal activity on C14 and C12 substrates. Further modification of the C12 enzyme yields a thioesterase having greater activity on C14 as compared to C12 substrates. Also provided in the instant invention are novel plant acyl-ACP thioesterase sequences from Cuphea palustris and mangosteen (Garcinia mangifera), which demonstrate desirable thioesterase activity profiles for plant genetic engineering applications.
    • 提供了改变植物酰基-ACP硫酯酶底物特异性的方法,以及如此制备的工程化植物酰基-ACP硫酯酶。 植物硫酯酶的C末端三分之二部分被鉴定为对于这种修饰是理想的。 还提供了用于表达工程化硫酯酶的DNA序列和构建体,以及由其产生的新的硫酯酶。 这样的DNA序列可用于在宿主细胞,特别是油籽作物植物的种子细胞中表达改性的硫酯酶,用于改变脂肪酸组成。 可以改变本文所述的C12优选植物酰基-ACP硫酯酶以获得对C14和C12底物具有大致相同活性的植物硫酯酶。 与C12底物相比,C12酶的进一步修饰产生对C14具有更大活性的硫酯酶。 本发明还提供了来自Cuphea palustris和山竹(Garcinia mangifera)的新型植物酰基-ACP硫酯酶序列,其显示用于植物基因工程应用的期望的硫酯酶活性谱。
    • 9. 发明授权
    • Anti-sense regulation of gene expression in plant cells
    • 植物细胞中基因表达的反义调节
    • US5107065A
    • 1992-04-21
    • US240408
    • 1988-08-30
    • Christine K. ShewmakerJean C. KridlWilliam R. HiattVic Knauf
    • Christine K. ShewmakerJean C. KridlWilliam R. HiattVic Knauf
    • C12N9/24C12N15/113C12N15/82
    • C12N15/1137C12N15/8216C12N15/8218C12N9/2402C12Y302/01015C12N2310/111
    • Regulation of expression of genes encoded for in plant cell genomes is achieved by integration of a gene under the transcriptional control of a promoter which is functional in the host and in which the transcribed strand of DNA is complementary to the strand of DNA that is transcribed from the endogenous gene(s) one wishes to regulate. The integrated gene, referred to as anti-sense, provides an RNA sequence capable of binding to naturally existing RNAs, exemplified by polygalacturonase, and inhibiting their expression, where the anti-sense sequence may bind to the coding, non-coding, or both, portions of the RNA. The antisense construction may be introduced into the plant cells in a variety of ways and be integrated into the plant genome for inducible or constitutive transcription of the anti-sense sequence. A wide variety of plant cell properties may be modifed by employing this technique.The pCGN978xK12 was deposited at the A.T.C.C. on Mar. 25, 1986, and given Accession No. 67064 and pCGN1401 was deposited on Oct. 7, 1986 at the A.T.C.C. and given Accession No. 67227.
    • 在植物细胞基因组中编码的基因的表达的调节是通过在宿主的功能的启动子的转录控制下整合基因来实现的,其中DNA的转录链与DNA转录的DNA互补 内源基因希望调节。 称为反义的整合基因提供了能够结合天然存在的RNA的RNA序列,例如由多聚半乳糖醛酸脱水酶,并抑制其表达,其中反义序列可以结合编码,非编码或两者 ,部分RNA。 反义结构可以以各种方式引入植物细胞中,并且整合到植物基因组中以诱导或组成型转录反义序列。 可以通过采用这种技术来改变各种各样的植物细胞特性。 pCGN978xK12保藏在A.T.C.C. 1986年3月25日,加拿大号67064和pCGN1401于1986年10月7日在A.T.C.C. 并提供登记号67227。