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1. 发明专利

JP2005185282A Method for obtaining circular mutated and/or chimeric polynucleotide 有权
标题翻译：获得圆形和/或CHIMERIC多核苷酸的方法
公开(公告)号：JP2005185282A
公开(公告)日：2005-07-14
申请号：JP2004350290
申请日：2004-12-02
申请人： F Hoffmann La Roche Ag , エフ.ホフマン−ラロシュアーゲーF. Hoffmann−La Roche Aktiengesellschaft
发明人： KRATZSCH PETER , VON DER ELTZ HERBERT , SCHMUCK RAINER , MARA BOENITZ-DULAT
IPC分类号： C12N15/09 , C12N1/15 , C12N1/19 , C12N1/21 , C12N5/10 , C12N9/16 , C12N15/10 , C12Q1/68
CPC分类号： C12N15/102 , C12N15/1027
摘要： PROBLEM TO BE SOLVED: To provide a method for simply and efficiently obtaining a chimeric polynucleotide by reducing process of operation while keeping high diversity of the chimeric nucleotide. SOLUTION: The invention relates to the method for generating the mutated and/or chimeric polynucleotide, comprising, (a) providing a closed circular polynucleotide comprising a first gene of the interest, (b) providing fragments of a double-stranded DNA target polynucleotide derived from a second gene of interest, capable of hybridizing to the first gene of interest, said fragments having free 3' OH ends and phosphorylated 5'-ends, (c) generating single strands of both said (a) and (b), (d) annealing said fragments of the target polynucleotide to said template polynucleotide, (e) elongating the annealed fragments of the target polynucleotide by DNA polymerase, (f) ligating the nicks between the elongated fragments by using a DNA ligase thereby generating a chimeric full length circular DNA molecule, and (g) using the product obtained in (f) and repeating steps (c) to (f) 1 to 100 times. COPYRIGHT: (C)2005,JPO&NCIPI
摘要翻译：待解决的问题：提供通过减少操作过程简单有效地获得嵌合多核苷酸的方法，同时保持嵌合核苷酸的高度多样性。解决方案：本发明涉及产生突变和/或嵌合多核苷酸的方法，包括（a）提供包含感兴趣的第一个基因的闭合环状多核苷酸，（b）提供双链DNA片段所述片段具有游离的3'OH末端和磷酸化的5'末端，（c）产生所述（a）和（b）的单链，），（d）将靶多聚核苷酸的所述片段退火至所述模板多核苷酸，（e）通过DNA聚合酶延伸靶多核苷酸的退火片段，（f）通过使用DNA连接酶连接细长片段之间的切口从而产生嵌合全长环状DNA分子，（g）使用（f）中得到的产物，重复步骤（c）〜（f）1〜100次。版权所有（C）2005，JPO＆NCIPI

2. 发明专利

JP2003180352A VARIANTS OF ERWINIA-TYPE CREATINASE 审中-公开
公开(公告)号：JP2003180352A
公开(公告)日：2003-07-02
申请号：JP2002275577
申请日：2002-09-20
申请人： HOFFMANN LA ROCHE
发明人： SHAO ZHIXIN , SCHMUCK RAINER , KRATZSCH PETER , KENKLIES JANET , WEISSER HARALD
IPC分类号： C12N15/09 , C12N1/21 , C12N9/78 , C12Q1/26 , C12Q1/28 , C12Q1/34 , C12R1/18
摘要： PROBLEM TO BE SOLVED: To provide a creatinase stably and efficiently usable in clinical and industrial application, and to provide their use for concentration measurement of creatinine and/or creatin in a sample. SOLUTION: The invention includes a modified Erwinia-type creatinase variant with creatinase activity comprising at lest one amino acid substitution at a selected position of a specific sequence; a reagent for measurement of creatinine containing the variant; a method of detection for creatin which incubates the objective sample with a reagent containing the variant, sarcosine oxidase, peroxidase and a reagent for detection of hydrogen peroxide, and then relates the product hydrogen peroxide with the concentration of the creatin in the sample; a method of detection for creatinine which incubates the objective sample with a reagent containing the variant, creatininase, sarcosine oxidase, peroxidase and a reagent for detection of hydrogen peroxide, and then relates the product hydrogen peroxide with the concentration of the creatinine in the sample. COPYRIGHT: (C)2003,JPO

3. 发明申请

WO03002736A3 A WALK-THROUGH TECHNIQUE FOR IN VITRO RECOMBINATION OF POLYNUCLEOTIDE SEQUENCES 审中-公开
标题翻译：用于多核苷酸序列的体外重组的WALK-THROUGH技术
公开(公告)号：WO03002736A3
公开(公告)日：2003-07-31
申请号：PCT/EP0207060
申请日：2002-06-26
申请人： ROCHE DIAGNOSTICS GMBH , HOFFMANN LA ROCHE , SHAO ZHIXIN , KRATZSCH PETER , SCHMUCK RAINER , ELTZ HERBERT VON DER , KENKLIES JANET
发明人： SHAO ZHIXIN , KRATZSCH PETER , SCHMUCK RAINER , VON DER ELTZ HERBERT , KENKLIES JANET
IPC分类号： C12N15/09 , C07K14/00 , C12N9/06 , C12N9/16 , C12N15/10 , C12P21/02
CPC分类号： C12N9/0034 , C12N9/16 , C12N15/102
摘要： A simple and efficient method for random in vitro mutagenesis and recombination of polynucleotide sequences based on walk-through with chain-elongating molecules and chain terminating molecules e.g. dNTPs/ddNTPs followed by reassembly and amplification is described. The utility of this method for improving protein structure and/or function is demonstrated by creating novel sarcosine oxidase variants and by recombining human placental alkaline phosphates and calf intestinal alkaline phosphatase for better mutants with improved properties.
摘要翻译：一种简单而有效的方法，用于随机体外诱变和基于穿越链延伸分子和链终止分子的多核苷酸序列的重组。描述了重组和扩增后的dNTPs / ddNTP。通过产生新的肌氨酸氧化酶变体和通过重组人胎盘碱性磷酸盐和小牛肠碱性磷酸酶来改善蛋白质结构和/或功能的这种方法来改善蛋白质结构和/或功能的更好的突变体具有改进的性质。

4. 发明专利

ES2803273T3 3-hidroxibutirato deshidrogenasa mutante de Alcaligenes faecalis, así como procedimientos y usos que implican la misma 未知
公开(公告)号：ES2803273T3
公开(公告)日：2021-01-25
申请号：ES16826037
申请日：2016-12-21
申请人： HOFFMANN LA ROCHE
发明人： KRATZSCH PETER , BOENITZ-DULAT MARA , BECK DANIELA , STREIDL THOMAS , HUNT DUVALL STACY
IPC分类号： C12N9/04
摘要： Una 3-hidroxibutirato deshidrogenasa (3-HBDH) mutante de Alcaligenes faecalis con un rendimiento mejorado en relación con la 3-HBDH natural, en la que la enzima mutante comprende una secuencia de aminoácidos que es al menos 80 % idéntica a la secuencia de aminoácidos de SEQ ID NO: 1 (3-HBDH de Alcaligenes faecalis; 3-HBDH natural) o la secuencia de aminoácidos de SEQ ID NO: 2 (secuencia central extendida de 3-HBDH natural) y en la que la enzima mutante tiene al menos tres sustituciones de aminoácidos en relación con la 3-HBDH natural, en la que el aminoácido en la posición correspondiente a - la posición 253 de SEQ ID NO: 1 se sustituye con Ile (253Ile), Ala (253Ala) o Cys (253Cys), - la posición 233 de SEQ ID NO: 1 se sustituye con Ser (233Ser), Ile (233Ile), Ala (233Ala), Pro (233Pro), Arg (233Arg), Thr (233Thr), Lys (233Lys) o Cys (233Cys), y - la posición 234 de SEQ ID NO: 1 se sustituye con Thr (234Thr).

5. 发明专利

CA2649135C IMPROVED MUTANTS OF PYRROLOQUINOLINE QUINONE DEPENDENT SOLUBLE GLUCOSE DEHYDROGENASE 未知
公开(公告)号：CA2649135C
公开(公告)日：2013-11-19
申请号：CA2649135
申请日：2007-04-11
申请人： HOFFMANN LA ROCHE
发明人： BOENITZ-DULAT MARA , BECK DANIELA , KRATZSCH PETER , SCHMUCK RAINER , VON DER ELTZ HERBERT
IPC分类号： C12N9/04
摘要： The present invention discloses a mutant of PQQ-dependent soluble glucose dehydrogenase (s-GDH; EC 1.1.5.2) with improved specificity for glucose as compared to maltose, having a substitution of threonine at position 348 by either glycine, alanine or serine, wherein said mutant additionally comprises, at least one mutation for improving the stability of the mutant and one or more mutation(s) for improving the affinity of the mutant to glucose, and/or one or more mutation(s) for further improving the specificity of the mutant for glucose as compared to maltose, and wherein position 348 correspond to the amino acid positions known from the A. calcoaceticuss-GDH wild-type sequence. Also disclosed are genes encoding such mutant s-GDH, and different applications of these s-GDH mutants, particularly for determining the concentration of glucose in a sample.

6. 发明专利

BRPI0710131A2 未知
公开(公告)号：BRPI0710131A2
公开(公告)日：2011-08-02
申请号：BRPI0710131
申请日：2007-04-11
申请人： HOFFMANN LA ROCHE
发明人： BOENITZ-DULAT MARA , BECK DANIELA , KRATZSCH PETER , SCHMUCK RAINER , VON DER ELTZ HERBERT
IPC分类号： C12N9/04
摘要： The present invention discloses a mutant of PQQ-dependent soluble glucose dehydrogenase (s-GDH; EC 1.1.5.2) with improved specificity for glucose as compared to maltose, having a substitution of threonine at position 348 by either glycine, alanine or serine, wherein said mutant additionally comprises, at least one mutation for improving the stability of the mutant and one or more mutation(s) for improving the affinity of the mutant to glucose, and/or one or more mutation(s) for further improving the specificity of the mutant for glucose as compared to maltose, and wherein position 348 correspond to the amino acid positions known from the A. calcoaceticus s-GDH wild-type sequence. Also disclosed are genes encoding such mutant s-GDH, and different applications of these s-GDH mutants, particularly for determining the concentration of glucose in a sample.

7. 发明专利

CA2427029C VARIANTS OF SOLUBLE PYRROLOQUINOLINE QUINONE-DEPENDENT GLUCOSE DEHYDROGENASE 未知
公开(公告)号：CA2427029C
公开(公告)日：2010-02-02
申请号：CA2427029
申请日：2001-10-20
申请人： HOFFMANN LA ROCHE
发明人： KNAPPE WOLFGANG-REINHOLD , SCHMUCK RAINER , KRATZSCH PETER , SHAO ZHIXIN , BECK DANIELA , THYM DETLEF
IPC分类号： C12N15/53 , G01N33/66 , C12M1/40 , C12N1/15 , C12N1/19 , C12N1/21 , C12N5/10 , C12N9/04 , C12N11/02 , C12N15/09 , C12P21/02 , C12Q1/32 , C12Q1/54
摘要： The present invention relates to improved variants of soluble pyrroloquinoli ne quinone (PQQ)-dependent glucose dehydrogenases (s-GDH), to genes encoding mutated s-GDH, to mutant proteins of s-GDH with improved substrate specifici ty for glucose, and to different applications of these s-GDH variants, particularly for determining concentrations of sugar, especially of glucose in a sample.

8. 发明专利

CA2453864A1 A WALK-THROUGH TECHNIQUE FOR IN VITRO RECOMBINATION OF POLYNUCLEOTIDE SEQUENCES 未知
公开(公告)号：CA2453864A1
公开(公告)日：2003-01-09
申请号：CA2453864
申请日：2002-06-26
申请人： HOFFMANN LA ROCHE
发明人： SCHMUCK RAINER , KENKLIES JANET , KRATZSCH PETER , SHAO ZHIXIN , VON DER ELTZ HERBERT
IPC分类号： C12N15/09 , C07K14/00 , C12N9/06 , C12N9/16 , C12N15/10 , C12P21/02
摘要： A simple and efficient method for random in vitro mutagenesis and recombination of polynucleotide sequences based on walk-through with chain- elongating molecules and chain terminating molecules e.g. dNTPs/ddNTPs followed by reassembly and amplification is described. The utility of this method for improving protein structure and/or function is demonstrated by creating novel sarcosine oxidase variants and by recombining human placental alkaline phosphates and calf intestinal alkaline phosphatase for better mutants with improved properties.

9. 发明专利

HUE049911T2 Pirrolokinolin-kinon-függõ, oldható glükóz-dehidrogenáz javított mutánsai 未知
公开(公告)号：HUE049911T2
公开(公告)日：2020-11-30
申请号：HUE17151869
申请日：2007-04-11
申请人： HOFFMANN LA ROCHE
发明人： BOENITZ-DULAT MARA , BECK DANIELA , KRATZSCH PETER , SCHMUCK RAINER , VON DER ELTZ HERBERT
IPC分类号： C12N9/04

10. 发明专利

ES2782199T3 3-Hidroxibutirato deshidrogenasa mutante de Rhodobacter sphaeroides, así como procedimientos y usos que implican la misma 未知
公开(公告)号：ES2782199T3
公开(公告)日：2020-09-11
申请号：ES17704245
申请日：2017-02-09
申请人： HOFFMANN LA ROCHE
发明人： BECK DANIELA , BOENITZ-DULAT MARA , KRATZSCH PETER , STREIDL THOMAS , HORN CARINA
IPC分类号： C12P7/18 , C12N9/04 , C12Q1/32
摘要： Una 3-hidroxibutirato deshidrogenasa (3-HBDH) mutante de Rhodobacter sphaeroides con un rendimiento mejorado en relación con la 3-HBDH natural en la que la enzima mutante comprende una secuencia de aminoácidos que es al menos un 80 % idéntica a la secuencia de aminoácidos de SEQ ID NO: 1 (3-HBDH de Rhodobacter sphaeroides; 3-HBDH natural) y en la que la enzima mutante tiene al menos una sustitución aminoacídica en relación con la 3-HBDH natural en una posición correspondiente a la posición 250 de SEQ ID NO: 1.

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