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    New restriction enzyme and process for producing the same
    2.
    发明授权
    New restriction enzyme and process for producing the same 失效
    新的限制酶及其制备方法

    公开(公告)号：US4808531A

    公开(公告)日：1989-02-28

    申请号：US756380

    申请日：1985-07-18

    申请人： Akira Obayashi ,  Nobutsugu Hiraoka ,  Keiko Kita ,  Hiroshi Nakajima

    发明人： Akira Obayashi ,  Nobutsugu Hiraoka ,  Keiko Kita ,  Hiroshi Nakajima

    IPC分类号： C12N15/09 ,  C12N9/16 ,  C12N9/22 ,  C12R1/01 ,  C12N9/14 ,  C12N9/00 ,  C12P21/00

    CPC分类号： C12N9/22 ,  Y10S435/822

    摘要： The invention provides restriction endonuclease Mf1 I capable of recognizing the base sequence as shown below on a double-stranded DNA molecule and cleaving the DNA chain at the arrow-marked positions, but has no such action when A is methylated5'--Pu.dwnarw.GATC Py--3'3'--Py CTAG.uparw.Pu--5'(wherein A represents adenosine, G guanosine, T thymidine, C cytidine, Pu adenosine or guanosine, and Py thymidine or cytidine). The restriction endonuclease is produced by culturing Microbacterium flavum IAM 1642, FERM BP-938 in a culture medium and recovering it from the culture.

    摘要翻译： 本发明提供了限制性内切核酸酶Mf1I，其能够识别如下所示的碱基序列在双链DNA分子上并且在箭头标记位置处切割DNA链，但当A是甲基化5'-Pu＆darr＆GATC时没有这样的作用 Py-3'3'-Py CTAG＆uarr＆Pu-5'（其中A表示腺苷，G鸟苷，T胸苷，C胞苷，Pu腺苷或鸟苷，Py胸苷或胞苷）。 限制性内切酶通过在培养基中培养黄色微生物IAM1642，FERM BP-938并从培养物中回收来产生。

    Process for producing restriction enzyme
    3.
    发明授权
    Process for producing restriction enzyme 失效
    生产限制酶的方法

    公开(公告)号：US4724209A

    公开(公告)日：1988-02-09

    申请号：US783634

    申请日：1985-10-03

    申请人： Akira Obayashi ,  Nobutsugu Hiraoka ,  Keiko Kita ,  Hiroshi Nakajima

    发明人： Akira Obayashi ,  Nobutsugu Hiraoka ,  Keiko Kita ,  Hiroshi Nakajima

    IPC分类号： C12N15/09 ,  C12N9/16 ,  C12N9/22 ,  C12R1/01 ,  C12N15/00 ,  C12Q1/68

    CPC分类号： C12N9/22 ,  Y10S435/822

    摘要： A process for producing a restriction endonuclease capable of recognizing the nucleotide sequence ##STR1## (wherein A, C, G and T represent adenosine, cytidine, guanosine and thymidine, respectively) on a DNA chain and specifically cleaving the double-stranded chain at the arrow-marked positions. This process comprises growing a microorganism belonging to the genus Halococcus and capable of producing said restriction endonuclease, and collecting the enzyme thus formed from the culture broth.

    摘要翻译： （其中A，C，G和T分别代表腺苷，胞苷，鸟苷和胸腺嘧啶核苷酸）分别限定在DNA链上并特异性切割双链的限制性内切核酸酶的方法 链在箭头标记的位置。 该方法包括生长属于卤虫属的微生物并且能够产生所述限制性内切核酸酶，并收集由培养液形成的酶。

    Novel restriction endonuclease SplI and process for the production of the same
    4.
    发明授权
    Novel restriction endonuclease SplI and process for the production of the same 失效
    新型限制性内切核酸酶SplI及其生产过程相同

    公开(公告)号：US4886756A

    公开(公告)日：1989-12-12

    申请号：US758536

    申请日：1985-07-24

    申请人： Masahide Kawamura ,  Masaki Sakakibara ,  Teruo Watanabe ,  Akira Obauashi ,  Nobutsugu Hiraoka ,  Keiko Kita

    发明人： Masahide Kawamura ,  Masaki Sakakibara ,  Teruo Watanabe ,  Akira Obauashi ,  Nobutsugu Hiraoka ,  Keiko Kita

    IPC分类号： C12N15/09 ,  C12N9/16 ,  C12N9/22 ,  C12R1/89

    CPC分类号： C12N9/22

    摘要： A novel restriction endonuclease SplI which has the following physicochemical properties:(1) recognizing the following base sequences in double-stranded deoxyribonucleic acid ##STR1## and cleaving said sequences in the phosphodiester bonds between C and G as indicated with the vertical arrows to produce DNA fragments having one strand comprising four bases at the 5'-terminal;(2) cleaving double-stranded deoxyribonucleic acid .lambda.-DNA in one position, Col El in two positions and .phi.x 174 RF in two positions;(3) being activated with 5 to 20 mM Mg.sup.2+ ; and(4) exhibiting an activity at a NaCl concentration of 0 to 200 mM;and a process for the production of the restriction endonuclease SplI which comprises culturing a restriction endonuclease SplI-producing alga belonging to the genus Spirulina, collecting the cells, obtaining a cell-free extract therefrom the separating and purifying the restriction endonuclease SplI.

    摘要翻译： 具有以下物理化学性质的新型限制性内切核酸酶SplI：（1）在双链脱氧核糖核酸中识别以下碱基序列，并用垂直箭头所示切割C和G之间的磷酸二酯键中的所述序列，以产生DNA 具有在5'末端具有4个碱基的一条链的片段; （2）在一个位置切割双链脱氧核糖核酸λ-DNA，两个位置分别为Col El，两个位置为174个RF; （3）用5〜20mM Mg 2+活化; 和（4）在NaCl浓度为0至200mM下表现出活性; 以及生产限制性内切核酸酶SplI的方法，其包括培养属于螺旋藻属的限制性内切核酸酶SplI产生藻类，收集细胞，从其中获得分离和纯化限制性内切核酸酶SplI的无细胞提取物。