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1. 发明申请

US20130267021A1 SYSTEM AND METHOD OF MODULAR CLONING 审中-公开
公开(公告)号：US20130267021A1
公开(公告)日：2013-10-10
申请号：US13703123
申请日：2011-06-09
申请人： Ernst Weber , Stefan Werner , Carola Engler , Ramona Grützner , Sylvestre Marillonnet
发明人： Ernst Weber , Stefan Werner , Carola Engler , Ramona Grützner , Sylvestre Marillonnet
IPC分类号： C12N15/66
CPC分类号： C12N15/66 , C12N15/1093
摘要： System for producing a nucleic acid construct of interest, said system comprising: a set of n entry DNAs numbered 1 to n, n being an integer of at least 2, each of said n entry DNAs comprising in this order: (i) a type IIs restriction endonuclease recognition site followed by the cleavage site thereof; (ii) a sequence portion linking the cleavage site of said recognition site of item (i) with the cleavage site of the recognition site of the following item (iii), and (iii) a cleavage site of a further type Ms restriction endonuclease recognition site followed by the recognition site of said cleavage site; the cleavage sites of the type IIs restriction endonuclease recognition sites of item (iii) of entry DNAs 1 to n−1 are complementary to the cleavage sites of the type IIs restriction endonuclease recognition sites of item (i) of entry DNAs 2 to n, respectively; the cleavage site of the type Ms restriction endonuclease recognition site of item (iii) of entry DNA n is complementary to the cleavage site of the type IIs restriction endonuclease recognition site of item (i) of entry DNA 1 for allowing annealing of complementary single-stranded overhangs formed by restriction at recognition site (i) of entry DNA 1 and at recognition site (iii) of entry DNA n; said system further comprising a destination vector comprising in this order: (I) a type Ms restriction endonuclease recognition site followed by the cleavage site thereof; (II) a vector backbone preferably comprising a selectable marker gene, said vector backbone linking the cleavage sites of said recognition sites of items (I) and the following item (III); (III) a further cleavage site of a type Ms restriction endonuclease recognition site followed by the recognition site of said cleavage site, and (IV) optionally, an insert between the recognition sites of item (III) and item (i); said cleavage sites of items (I) and (III) being different and non-complementary, said recognition sites of items (I) and (III) being preferably recognitions sites of the same endonuclease.

2. 发明授权

US08883420B2 Process of clean cloning 有权
标题翻译：清洁克隆的过程
公开(公告)号：US08883420B2
公开(公告)日：2014-11-11
申请号：US13121019
申请日：2009-10-08
申请人： Sylvestre Marillonnet , Stefan Werner , Carola Engler , Romy Kandzia , Frank Thieme , Ernst Weber
发明人： Sylvestre Marillonnet , Stefan Werner , Carola Engler , Romy Kandzia , Frank Thieme , Ernst Weber
IPC分类号： C12Q1/68 , C12N15/66 , C12N15/64
CPC分类号： C12N15/64 , C12N15/66
摘要： A process of inserting a nucleic acid sequence of interest into an acceptor nucleic acid is provided. The process comprises amplifying by PCR a DNA comprising in the following order a sequence segment U, a nucleic acid sequence segment of known nucleotide sequence K2, and a nucleic acid sequence segment of known sequence K3. The process further comprises treating the linear double-stranded DNA molecules from the PCR amplification with an exonuclease to obtain a single-stranded overhang at the first end of the DNA and a single-stranded overhang comprising nucleic acid segments K2 and K3 at the second end of the DNA. The process additionally comprises annealing the product of the exonuclease treatment to a linearized double-stranded acceptor nucleic acid which has been designed to complement the single-stranded overhangs of the product of the exonuclease treatment.
摘要翻译：提供将感兴趣的核酸序列插入受体核酸的过程。该方法包括通过PCR扩增包含以下顺序的序列片段U，已知核苷酸序列K2的核酸序列片段和已知序列K3的核酸序列片段的DNA。该方法还包括用外切核酸酶从PCR扩增中处理线性双链DNA分子，以在DNA的第一端获得单链突出端，并且在第二末端包含包含核酸片段K2和K3的单链突出端的DNA。该方法还包括将外切核酸酶处理的产物退火至线性化双链受体核酸，其被设计为补充外切核酸酶处理产物的单链突出端。

3. 发明申请

US20110263024A1 PROCESS OF CLEAN CLONING 有权
标题翻译：清洁过程
公开(公告)号：US20110263024A1
公开(公告)日：2011-10-27
申请号：US13121019
申请日：2009-10-08
申请人： Sylvestre Marillonnet , Stefan Werner , Carola Engler , Romy Kandzia , Frank Thieme , Ernst Weber
发明人： Sylvestre Marillonnet , Stefan Werner , Carola Engler , Romy Kandzia , Frank Thieme , Ernst Weber
IPC分类号： C12N15/87
CPC分类号： C12N15/64 , C12N15/66
摘要： A process of inserting a nucleic acid sequence of interest into an acceptor nucleic acid, comprising the following steps: amplifying by PCR a DNA comprising in the following order a sequence segment U, a nucleic acid sequence segment of known nucleotide sequence K2 and a nucleic acid sequence segment of known sequence K3 using a forward primer defining a first end of the amplified DNA and a reverse primer defining a second end of the amplified DNA, said reverse primer terminating at its 3′-end in a nucleotide sequence of nucleic acid sequence segment K3; treating the linear double-stranded DNA molecules contained in the PCR product obtained in the previous step with an exonuclease to obtain a single-stranded overhang at the first end of the DNA and a single-stranded overhang comprising nucleic acid segments K2 and K3 at the second end of the DNA; and annealing the product of the previous step to a linearized double-stranded acceptor nucleic acid having at a first end thereof a single-stranded overhang complementary to the single-stranded overhang of the first end of the DNA and at a second end thereof a single-stranded overhang complementary to the single-stranded sequence segment K2 of the second end of the DNA.
摘要翻译：包括以下步骤：通过PCR扩增包含以下顺序的DNA序列的已知核苷酸序列K2的核酸序列片段和核酸的方法已知序列K3的序列片段使用限定扩增DNA第一末端的正向引物和限定扩增DNA第二末端的反向引物，所述反向引物在其核酸序列片段的核苷酸序列的3'末端终止 K3; 用外切核酸酶处理前一步骤中获得的PCR产物中包含的线性双链DNA分子，以在DNA的第一端获得单链突出端，并且在第一端包含核酸片段K2和K3的单链突出端 DNA的第二端; 并将前述步骤的产物退火到线性双链受体核酸，其在第一端具有与DNA的第一末端的单链突出部互补的单链突出端，并且在其第二端具有单链突出端与DNA的第二末端的单链序列片段K2互补的突出突出端。

4. 发明授权

US4735014A Method of and apparatus for industrial cultivation, particularly of mushrooms and also the substrate used for such cultivation 失效
标题翻译：用于工业栽培的方法和设备，特别是蘑菇以及用于这种栽培的底物
公开(公告)号：US4735014A
公开(公告)日：1988-04-05
申请号：US864793
申请日：1986-05-19
申请人： Ernst Weber
发明人： Ernst Weber
IPC分类号： A01D45/00 , A01G1/04
CPC分类号： A01D45/005 , A01G1/042
摘要： (a) The invention relates to a method of and an apparatus for industrially growing particularly mushrooms and also to a substrate used for such cultivation.(b) A method characterized in that a substrate is used which consists of a mixture based on hemp stalks, the substrate being prepared by impregnation in water, whereupon the substrate is fixed on supports constituting vertical walls, whereupon the apparatus (1) carrying the substrate is placed in an incubation room and then in a growing room.(c) The present invention has its main application in the mushrooms growing industry.
摘要翻译：（a）本发明涉及特别是蘑菇的工业生长方法和装置，还涉及用于这种栽培的底物。（b）一种方法，其特征在于，使用由基于大麻茎的混合物组成的基材，所述基材通过在水中浸渍制备，然后将基材固定在构成垂直壁的支撑体上，于是装载（1）将底物放置在孵育室中，然后放置在生长室中。（c）本发明主要应用于蘑菇种植业。

5. 发明授权

US4927544A Process for the continuous removal of a gum phase from triglyceride oil 失效
标题翻译：从甘油三酯油连续除去胶相的方法
公开(公告)号：US4927544A
公开(公告)日：1990-05-22
申请号：US322860
申请日：1989-03-14
申请人： Martin van Opstal , Frans Dewulf , Bernard Cleenewerck , Ernst Weber
发明人： Martin van Opstal , Frans Dewulf , Bernard Cleenewerck , Ernst Weber
IPC分类号： C11B3/00 , C11B3/02 , C11B3/16 , B01D21/26
CPC分类号： C11B3/16 , C11B3/001 , C11B3/02
摘要： A three or optionally four stage process for the continuous removal of a gum phase from triglyceride oil is described in which in a first stage the oil containing a separate gum phase is subjected to centrifugal separation in a first centrifugal separator to yield gums with low oil content and an oil that still contains a fraction of the gums originally present in the feed, in a second stage the oil obtained from the first stage is subjected to centrifugal separation in a second centrifugal separator to yield oil with a further reduced residual gum content and a gum phase with a higher oil content than the gums obtained in the first stage, in a third stage the gum phase obtained in the second stage is recycled into the oil stream fed to the first centrifugal separator and optionally in a fourth stage the oil obtained in the second stage is washed one or more times with water.

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