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1. 发明申请

WO2006093529A3 COVALENT TETHERING OF FUNCTIONAL GROUPS TO PROTEINS AND SUBSTRATES THEREFOR 审中-公开
标题翻译：功能组合对蛋白质和底物的共价分析
公开(公告)号：WO2006093529A3
公开(公告)日：2007-03-22
申请号：PCT/US2005027307
申请日：2005-07-29
申请人： PROMEGA CORP , DARZINS ALDIS , ENCELL LANCE P , JOHNSON TONNY , KLAUBERT DIETER , LOS GEORGYI V , MCDOUGALL MARK , WOOD KEITH V , WOOD MONIKA G , ZIMPRICH CHAD
发明人： DARZINS ALDIS , ENCELL LANCE P , JOHNSON TONNY , KLAUBERT DIETER , LOS GEORGYI V , MCDOUGALL MARK , WOOD KEITH V , WOOD MONIKA G , ZIMPRICH CHAD
IPC分类号： G01N33/58 , C07K14/47 , C12N9/14 , C12Q1/26 , C12Q1/34
CPC分类号： C12N9/14 , B82Y5/00 , B82Y10/00 , B82Y30/00 , C07K2299/00 , C12N9/0069 , C12Q1/26 , C12Q1/34 , C12Q1/66 , C12Y113/12005 , C12Y308/01005
摘要： A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.
摘要翻译：提供任选融合到感兴趣的蛋白质的突变体水解酶。突变体水解酶能够与相应的非突变（野生型）水解酶的底物形成键，其比野生型水解酶和底物之间形成的键更稳定，并且相对于野生型水解酶具有至少两个氨基酸取代野生型水解酶。还提供了包含一个或多个官能团的水解酶的底物，以及使用本发明的突变体水解酶和底物的方法。还提供能够与底物形成稳定结合的融合蛋白和表达融合蛋白的细胞。

2. 发明申请

WO2008054821A3 MUTANT HYDROLASE PROTEINS WITH ENHANCED KINETICS AND FUNCTIONAL EXPRESSION 审中-公开
标题翻译：具有增强动力学和功能表达的突变型水解蛋白
公开(公告)号：WO2008054821A3
公开(公告)日：2008-12-24
申请号：PCT/US2007023205
申请日：2007-10-30
申请人： PROMEGA CORP , DARZINS ALDIS , ENCELL LANCE P , OHANA RACHEL FRIEDMAN , OTTO PAUL , VIDUGIRIS GEDIMINAS , WOOD KEITH V , WOOD MONIKA G , ZIMMERMAN KRIS , SLATER MICHAEL R , ZHAO KATE QIN
发明人： DARZINS ALDIS , ENCELL LANCE P , OHANA RACHEL FRIEDMAN , OTTO PAUL , VIDUGIRIS GEDIMINAS , WOOD KEITH V , WOOD MONIKA G , ZIMMERMAN KRIS , SLATER MICHAEL R , ZHAO KATE QIN
IPC分类号： C12N9/14 , C12Q1/34
CPC分类号： C12N9/14 , C12N9/96 , C12N11/00 , C12Q1/34 , C12Y308/01 , G01N33/581
摘要： The invention provides a mutant hydrolase protein with enhanced kinetics and functional expression, as well as polynucleotides encoding the mutant proteins and methods of using the polynucleotides and mutant proteins.
摘要翻译：本发明提供具有增强的动力学和功能表达的突变水解酶蛋白，以及编码突变蛋白的多核苷酸以及使用多核苷酸和突变蛋白的方法。

3. 发明申请

WO2008054821A2 MUTANT HYDROLASE PROTEINS WITH ENHANCED KINETICS AND FUNCTIONAL EXPRESSION 审中-公开
标题翻译：突变体水解酶蛋白具有增强的动力学和功能表达
公开(公告)号：WO2008054821A2
公开(公告)日：2008-05-08
申请号：PCT/US2007/023205
申请日：2007-10-30
申请人： PROMEGA CORPORATION , DARZINS, Aldis , ENCELL, Lance, P. , OHANA, Rachel, Friedman , OTTO, Paul , VIDUGIRIS, Gediminas , WOOD, Keith, V. , WOOD, Monika, G. , ZIMMERMAN, Kris , SLATER, Michael, R. , ZHAO, Kate, Qin
发明人： DARZINS, Aldis , ENCELL, Lance, P. , OHANA, Rachel, Friedman , OTTO, Paul , VIDUGIRIS, Gediminas , WOOD, Keith, V. , WOOD, Monika, G. , ZIMMERMAN, Kris , SLATER, Michael, R. , ZHAO, Kate, Qin
IPC分类号： C12N9/14
CPC分类号： C12N9/14 , C12N9/96 , C12N11/00 , C12Q1/34 , C12Y308/01 , G01N33/581
摘要： The invention provides a mutant hydrolase protein with enhanced kinetics and functional expression, as well as polynucleotides encoding the mutant proteins and methods of using the polynucleotides and mutant proteins.
摘要翻译：本发明提供了具有增强的动力学和功能表达的突变水解酶蛋白，以及编码突变蛋白的多核苷酸以及使用多核苷酸和突变蛋白的方法。

4. 发明申请

WO2004015084A3 MUTANT E. COLI APPA PHYTASE ENZYMES 审中-公开
标题翻译： MUTANT E.CEL APPA PHYTASE ENZYMES
公开(公告)号：WO2004015084A3
公开(公告)日：2004-10-28
申请号：PCT/US0325058
申请日：2003-08-11
申请人： GENENCOR INT , BLATTMANN BEAT O , DARZINS ALDIS , DAVIS JOHN M , ENCELL LANCE P , MORRISON THOMAS B , MRACHKO GREGORY T , SCHELLENBERGER VOLKER
发明人： BLATTMANN BEAT O , DARZINS ALDIS , DAVIS JOHN M , ENCELL LANCE P , MORRISON THOMAS B , MRACHKO GREGORY T , SCHELLENBERGER VOLKER
IPC分类号： C12N9/16 , C12N15/15
CPC分类号： C12N15/8243 , C12N9/16 , C12N15/8245
摘要： Cells producing mutant phytases having modified activity are provided, as well as the phytases so produced. Also provided are methods of making and producing such phytases and the use of the expressed phytase protein in feed as a supplement.
摘要翻译：提供具有改变活性的产生突变体植酸酶的细胞，以及如此制备的植酸酶。还提供了制备和生产这种植酸酶的方法以及在饲料中使用表达的植酸酶蛋白作为补充物。

5. 发明申请

WO2004015084A2 MUTANT E. COLI APPA PHYTASE ENZYMES AND NATURAL VARIANTS THEREOF, NUCLEIC ACIDS ENCODING SUCH PHYTASE ENZYMES, VECTORS AND HOST CELLS INCORPORATING SAME AND METHODS OF MAKING AND USING SAME 审中-公开
标题翻译：突变体E.coli APPA PHYTASE ENZYMES及其自然变体，编码与其相同的这种植物酶，载体和宿主细胞的核酸及其制备和使用方法
公开(公告)号：WO2004015084A2
公开(公告)日：2004-02-19
申请号：PCT/US2003/025058
申请日：2003-08-11
申请人： GENENCOR INTERNATIONAL, INC. , BLATTMANN, Beat, O. , DARZINS, Aldis , DAVIS, John, M. , ENCELL, Lance, P. , MORRISON, Thomas, B. , MRACHKO, Gregory, T. , SCHELLENBERGER, Volker
发明人： BLATTMANN, Beat, O. , DARZINS, Aldis , DAVIS, John, M. , ENCELL, Lance, P. , MORRISON, Thomas, B. , MRACHKO, Gregory, T. , SCHELLENBERGER, Volker
IPC分类号： C12N
CPC分类号： C12N15/8243 , C12N9/16 , C12N15/8245
摘要： Cells producing mutant phytases having modified activity are provided, as well as the phytases so produced. Also provided are methods of making and producing such phytases and the use of the expressed phytase protein in feed as a supplement.
摘要翻译：提供具有改变活性的产生突变体植酸酶的细胞，以及如此制备的植酸酶。还提供了制备和生产这种植酸酶的方法以及在饲料中使用表达的植酸酶蛋白作为补充物。

6. 发明申请

WO0129211A9 METHOD FOR DIRECTED EVOLUTION BY RANDOM CHIMERAGENESIS ON TRANSIENT TEMPLATES 审中-公开
标题翻译：通过随机氯化物对瞬态模板进行定向演化的方法
公开(公告)号：WO0129211A9
公开(公告)日：2002-08-15
申请号：PCT/US0029046
申请日：2000-10-19
申请人： ENCHIRA BIOTECHNOLOGY CORP , COCO WAYNE M , ARENSDORF JOSEPH J , CRIST MICHAEL J , DARZINS ALDIS , ENCELL LANCE P , FOLSOM BRIAN R , HEKTOR HANS J , NALTY MARK S , LEVINSON WILLIAM E , PALMER STEPHEN R
发明人： COCO WAYNE M , ARENSDORF JOSEPH J , CRIST MICHAEL J , DARZINS ALDIS , ENCELL LANCE P , FOLSOM BRIAN R , HEKTOR HANS J , NALTY MARK S , LEVINSON WILLIAM E , PALMER STEPHEN R
IPC分类号： C12N15/10 , C12P19/34 , C12Q1/68
CPC分类号： C12N15/1031 , C12N15/1027 , C12P19/34
摘要： A method is provided wherein at least one template is used to align oligonucleotides of interest or fragments thereof to generate chimeric polynucleotides. The oligonucleotides are allowed to hybridize to the template or templates. If necessary, unhybridized termini of hybridized oligonucleotides are removed and gaps between hybridized oligonucleotides are filled. Adjacently hybridized oligonucleotides are ligated to generate chimeric polynucleotides hybridized to the template or templates. The chimeric polynucleotides can then be subjected to selection and/or screening for desired characteristics. Prior to or concomitant with selection/screening, the template is optionally removed.
摘要翻译：提供了一种方法，其中使用至少一个模板来配对目的寡核苷酸或其片段以产生嵌合多核苷酸。允许寡核苷酸与模板或模板杂交。如果需要，杂交的寡核苷酸的未杂交的末端被去除并且杂交的寡核苷酸之间的间隙被填充。连接相邻杂交的寡核苷酸以产生与模板或模板杂交的嵌合多核苷酸。然后可以对嵌合多核苷酸进行选择和/或筛选所需的特征。在选择/筛选之前或伴随选择/筛选，任选地去除模板。

7. 发明申请

WO0129211A3 METHOD FOR DIRECTED EVOLUTION BY RANDOM CHIMERAGENESIS ON TRANSIENT TEMPLATES 审中-公开
标题翻译：用瞬态模式对瞬态模板进行有向演化的方法
公开(公告)号：WO0129211A3
公开(公告)日：2002-01-24
申请号：PCT/US0029046
申请日：2000-10-19
申请人： ENCHIRA BIOTECHNOLOGY CORP , COCO WAYNE M , ARENSDORF JOSEPH J , CRIST MICHAEL J , DARZINS ALDIS , ENCELL LANCE P , FOLSOM BRIAN R , HEKTOR HANS J , NALTY MARK S , LEVINSON WILLIAM E , PALMER STEPHEN R
发明人： COCO WAYNE M , ARENSDORF JOSEPH J , CRIST MICHAEL J , DARZINS ALDIS , ENCELL LANCE P , FOLSOM BRIAN R , HEKTOR HANS J , NALTY MARK S , LEVINSON WILLIAM E , PALMER STEPHEN R
IPC分类号： C12N15/10 , C12P19/34 , C12Q1/68
CPC分类号： C12N15/1031 , C12N15/1027 , C12P19/34
摘要： A method is provided wherein at least one template is used to align oligonucleotides of interest or fragments thereof to generate chimeric polynucleotides. The oligonucleotides are allowed to hybridize to the template or templates. If necessary, unhybridized termini of hybridized oligonucleotides are removed and gaps between hybridized oligonucleotides are filled. Adjacently hybridized oligonucleotides are ligated to generate chimeric polynucleotides hybridized to the template or templates. The chimeric polynucleotides can then be subjected to selection and/or screening for desired characteristics. Prior to or concomitant with selection/screening, the template is optionally removed.
摘要翻译：提供了一种方法，其中至少一个模板用于比对目标寡核苷酸或其片段以产生嵌合多核苷酸。允许寡核苷酸与模板或模板杂交。如果需要，去除杂交寡核苷酸的未杂交末端并填充杂交寡核苷酸之间的空位。连接相邻杂交的寡核苷酸以产生与模板或模板杂交的嵌合多核苷酸。然后可以对嵌合多核苷酸进行选择和/或筛选期望的特征。在选择/筛选之前或伴随选择/筛选，模板可选地被移除。

8. 发明申请

WO1996040943A1 USE OF GRAM-POSITIVE BACTERIA TO EXPRESS RECOMBINANT PROTEINS 审中-公开
标题翻译：使用GRAM阳性细菌来表达重组蛋白
公开(公告)号：WO1996040943A1
公开(公告)日：1996-12-19
申请号：PCT/US1996009965
申请日：1996-06-06
申请人： THE ROCKEFELLER UNIVERSITY , DARZINS, Aldis , WHITEHEAD, Stephen , HRUBY, Dennis
发明人： THE ROCKEFELLER UNIVERSITY
IPC分类号： C12N15/62
CPC分类号： C07K14/31 , A61K38/00 , A61K39/00 , C07K14/315 , C07K2319/00 , C07K2319/02 , C07K2319/035 , C07K2319/23 , C07K2319/50 , C07K2319/705 , C07K2319/735 , C12N15/62 , C12N15/74
摘要： A novel system for cloning and expression of genes in gram-positive bacteria. The expression system is based on the finding that many gram-positive bacteria sort proteins to their cell surface through cis-acting N-terminal signal sequences and C-terminal anchor regions. In particular, the cell sorting signals of the streptococcal M6 protein, a well-known surface molecule, are used to construct a gram-positive expression system, designated SPEX (Streptococcal Protein Expression). Expression is achieved by cloning the gene of interest into an appropriate SPEX cassette which is then stably introduced into a bacterial host, such as the human commensal Streptococcus gordonii. Depending on the SPEX vector used, recombinant proteins can be anchored to the cell wall prior to release by specific endoproteolytic cleavage or secreted into the culture medium during bacterial growth. The use of host bacteria lacking extracellular proteases should protect secreted proteins from proteolytic degradation. Several expression vectors in this system also produce specifically-tagged recombinant proteins which allows for a one-step purification of the resulting product.
摘要翻译：用于克隆和表达革兰氏阳性菌中基因的新系统。表达系统基于许多革兰氏阳性细菌通过顺式作用的N末端信号序列和C末端锚定区域将蛋白质分解到其细胞表面的发现。特别地，使用众所周知的表面分子的链球菌M6蛋白的细胞分选信号来构建称为SPEX（链球菌蛋白表达）的革兰氏阳性表达系统。通过将感兴趣的基因克隆到适当的SPEX盒中，然后将其稳定地导入细菌宿主例如人共生型链球菌（Streptococcus gordonii）中来实现表达。根据所使用的SPEX载体，重组蛋白质可以在通过特异性内切蛋白酶切割释放之前锚定到细胞壁上，或在细菌生长过程中分泌到培养基中。缺乏细胞外蛋白酶的宿主细菌的使用应保护分泌的蛋白质免于蛋白水解降解。该系统中的几种表达载体也产生特异性标记的重组蛋白，其允许对所得产物进行一步纯化。

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