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    • 3. 发明申请
    • ALKALINE PROTEASE, PROCESS FOR THE PRODUCTION THEREOF, USE THEREOF, AND MICROORGANISM PRODUCING THE SAME
    • 碱性蛋白酶,其生产方法,用途及其生产的微生物
    • WO1997016541A1
    • 1997-05-09
    • PCT/JP1996003216
    • 1996-11-01
    • NOVO NORDISK A/SMIYOTA, YoshiakiFUKUYAMA, ShiroYONEDA, Tadashi
    • NOVO NORDISK A/S
    • C12N09/54
    • C12R1/07C11D3/386C11D3/38609C11D3/38618C12N9/54
    • A novel strain of Bacillus sp. (accession number: FERM BP-5736) and a protease which is obtained by incubating this strain or its variant, shows a high stability in the presence of surfactants, and has the following properties: (1) hydrolyzing proteins and peptides; (2) having an optimum pH value of around 12, when reacted with the use of casein as the substrate at 30 DEG C for 10 minutes; (3) having a stable pH value of from 5 to 11, when incubated with the use of casein as the substrate at 30 DEG C for 24 hours; (4) having an optimum action temperature of around 60 DEG C, when reacted with the use of casein as the substrate at pH 10 for 10 minutes; (5) having a molecular weight of 29,000 +/- 2,000 determined by SDS polyacrylamide gel electrophoresis; (6) having an isoelectric piont of 10.1 +/- 0.5 determined by isoelectric polyacrylamide gel electrphoresis; and (7) having a residual activity of 60 % or above in a 500 ppm LAS solution after 30 minutes at 40 DEG C or 20 % or above after 15 minutes at 55 DEG C.
    • 一种新型芽孢杆菌菌株 (登录号:FERM BP-5736)和通过温育该菌株或其变体获得的蛋白酶在表面活性剂存在下显示出高稳定性,具有以下性质:(1)水解蛋白质和肽; (2)当在30℃下与酪蛋白作为底物反应10分钟时,具有约12的最佳pH值; (3)当在30℃下使用酪蛋白作为底物孵育24小时时,其稳定的pH值为5至11; (4)在pH 10下与酪蛋白作为底物反应10分钟时,具有约60℃的最佳作用温度; (5)通过SDS聚丙烯酰胺凝胶电泳测定分子量为29,000 +/- 2,000; (6)通过等电聚丙烯酰胺凝胶电泳测定等电点为10.1 +/- 0.5; 和(7)在55℃下在30分钟后在30分钟内在500ppm LAS溶液中具有60%或更高的残留活性,在55℃下15分钟后具有20%或更高的残余活性。
    • 4. 发明申请
    • DOUBLE-STRANDED DNA WITH COHESIVE END(S), AND METHOD OF SHUFFLING DNA USING THE SAME
    • 具有粘结末端的双链DNA和使用其的DNA的方法
    • WO1998005765A1
    • 1998-02-12
    • PCT/DK1997000317
    • 1997-07-23
    • NOVO NORDISK A/SMIYOTA, YoshiakiFUKUYAMA, Shiro
    • NOVO NORDISK A/S
    • C12N15/10
    • C12N15/10C12N15/1027C12N15/66C12Q1/68C12Q1/6811
    • To provide a method of mutation of DNAs, which is substantially different from the conventional methods applicable to naturally-existing DNAs, and also to provide useful genetic products to be produced by the use of thus-mutated DNAs. A DNA with a cohesive end comprising (a) a double-stranded DNA having the same sequence as that of a part of a gene, and (b) a single-stranded DNA having a base sequence that exists on said gene at the site not adjoining the part corresponding to said double-stranded DNA or a base sequence which said gene does not have, wherein the single-stranded DNA is linked to either one end of the double-stranded DNA to form a cohesive end; a method for producing it; a method of shuffling a DNA using it; a DNA and a DNA pool to be obtained by the shuffling method; a method for producing the DNA pool; and a genetic product to be obtained by expressing the genetic information existing in the DNA pool.
    • 提供DNA的突变方法,其与适用于天然存在的DNA的常规方法基本上不同,并且还提供通过使用由此突变的DNA产生的有用的遗传产物。 具有粘性末端的DNA包含(a)与基因的一部分具有相同序列的双链DNA,和(b)在所述位点处存在于所述基因上的碱基序列的单链DNA, 邻接对应于所述双链DNA的部分或所述基因不具有的碱基序列,其中所述单链DNA连接到所述双链DNA的任一端以形成粘性末端; 一种生产它的方法; 使用它洗脱DNA的方法; 通过洗牌法获得的DNA和DNA库; 一种生产DNA池的方法; 以及通过表达DNA池中存在的遗传信息而获得的遗传产物。