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1. 发明申请

US20150031085A1 Novel Leech Hyaluronidase and Its Application 有权
标题翻译：新型水蛭透明质酸酶及其应用
公开(公告)号：US20150031085A1
公开(公告)日：2015-01-29
申请号：US14251579
申请日：2014-04-12
申请人： Jian Chen , Guocheng Du , Zhen Kang , Peng Jin
发明人： Jian Chen , Guocheng Du , Zhen Kang , Peng Jin
IPC分类号： C12N9/24 , C12P19/26
CPC分类号： C12N9/2402 , C12P19/26 , C12P21/02 , C12Y302/01036
摘要： The present invention provides a novel leech HAase and a method of producing low-molecular-weight HA oligosaccharides using the leech HAase. This invention successfully cloned the first leech HAase gene and provides a method for high-level expression of the leech HAase gene. By controlling the incubation condition, different HA oligosaccharides, particularly HA4, HA6, HA8 and HA10, can be selectively generated using the leech HAase. The large-scale expression of the leech HAase and the enzymatic production of specific HA oligosaccharides are not only useful for the cosmetic, healthcare and the medical industries but also can be a great help to polysaccharides chemical synthesis and cancer research.
摘要翻译：本发明提供了一种新的水蛭HAase和使用该水蛭HAase生产低分子量HA寡糖的方法。本发明成功克隆了第一个水蛭HAase基因，提供了一种高水平表达水蛭HAase基因的方法。通过控制孵育条件，可以使用水蛭HAase选择性地产生不同的HA寡糖，特别是HA4，HA6，HA8和HA10。水蛭HAase的大规模表达和特异性HA寡糖的酶促生产不仅可用于化妆品，保健和医疗行业，而且可以有助于多糖化学合成和癌症研究。

2. 发明申请

US20160319272A1 Method for Directed DNA Evolution using Combinatorial DNA Libraries 审中-公开
标题翻译：使用组合DNA文库进行定向DNA进化的方法
公开(公告)号：US20160319272A1
公开(公告)日：2016-11-03
申请号：US15140408
申请日：2016-04-27
申请人： Zhen Kang , Jian Chen , Guocheng Du , Peng Jin
发明人： Zhen Kang , Jian Chen , Guocheng Du , Peng Jin
IPC分类号： C12N15/10
CPC分类号： C12N15/1058 , C12N15/1031 , C12Q2521/101 , C12Q2521/501 , C12Q2531/113 , C12Q2533/101 , C12Q2563/179
摘要： The present invention provides a method of rapid directed DNA evolution based on single-stranded combinatorial DNA mutant library. The single- and double-stranded mutant library are constructed either separately or simultaneously using editing primers that contain mutated nucleotides and are targeted to different regions of the parent DNA sequence. The mutant library is then inserted into expression vectors and mutants with desired property are obtained by high throughput screening. Evolution of promoters, enzymes and metabolic pathways were successfully achieved using this method and mutants with excellent properties were obtained. The method of the present invention is simple, rapid, and efficient. It can be used for directed evolution of regulatory sequence such as promoters and ribosome binding sites, and is especially suitable for introducing diverse mutations into protein encoding genes, leading to rapid directed evolution of gene of interest.
摘要翻译：本发明提供了基于单链组合DNA突变体文库的快速定向DNA进化的方法。使用含有突变核苷酸的编辑引物单独或同时构建单链和双链突变体文库，并且靶向母DNA序列的不同区域。然后将突变型文库插入表达载体中，并通过高通量筛选获得具有所需性质的突变体。使用该方法成功实现了启动子，酶和代谢途径的进化，获得了具有优异性能的突变体。本发明的方法简单，快速，高效。它可以用于调节序列的定向进化，如启动子和核糖体结合位点，特别适用于将多种突变引入到蛋白质编码基因中，导致感兴趣的基因的快速定向进化。

3. 发明授权

US10793888B2 In vitro method for fast scarless DNA assembly using thermostable exonucleases and ligase 有权
公开(公告)号：US10793888B2
公开(公告)日：2020-10-06
申请号：US15347750
申请日：2016-11-09
申请人： Zhen Kang , Jian Chen , Peng Jin , Guocheng Du , Wenwen Ding
发明人： Zhen Kang , Jian Chen , Peng Jin , Guocheng Du , Wenwen Ding
IPC分类号： C12P19/34 , C12Q1/6806 , C12N15/10
摘要： The present invention provides a method for scarless in vitro DNA assembly using thermostable exonucleases and ligase, which relates to the field of genetic engineering. The present invention provides a fast method for assembling DNA subfragments with homologous ends, which employs thermostable polymerases and ligase in a thermal cycle of denaturation, annealing, digestion and ligation. After denaturation, DNA subfragments are assembled together via annealing of the homologous end sequences, the unpaired single-stranded overhangs are digested by polymerases, and the resulting nicked gaps are sealed by a ligase. Using this method, 2-6 DNA subfragments were successfully assembled within two hours. This method can be used in conventional DNA recombination and be adapted to high throughput assembly operations. In addition, combinatorial mutations can be easily introduced into the assembled sequence by use of primers with mutated bases. It is particularly suitable for making enzyme and synthetic pathways mutation libraries with high diversity, which can be used in directed evolution to screen for enzymes and synthetic pathways with desirable properties.

4. 发明授权

US09771607B2 Method of constructing a recombinant Bacillus subtilis that can produce specific-molecular-weight hyaluronic acids 有权
公开(公告)号：US09771607B2
公开(公告)日：2017-09-26
申请号：US15011509
申请日：2016-01-30
申请人： Jian Chen , Zhen Kang , Guocheng Du , Peng Jin
发明人： Jian Chen , Zhen Kang , Guocheng Du , Peng Jin
IPC分类号： C07K14/00 , C12P19/26 , C12N9/26 , C12N9/04 , C12N9/12 , C12N9/90 , C12N9/10
CPC分类号： C12P19/26 , C12N9/0006 , C12N9/1096 , C12N9/1241 , C12N9/2474 , C12N9/90 , C12Y101/01022 , C12Y206/01016 , C12Y207/07009 , C12Y207/07023 , C12Y302/01035 , C12Y504/0201
摘要： The present invention relates to the field of biotechnology engineering. It provides a method of constructing a recombinant Bacillus subtilis that can produce specific-molecular-weight hyaluronic acids. By integranted expression of hasA from Streptococcus zooepidemicus and overexpression of genes of HA synthetic pathway, tuaD, glmU and glmS, high yield HA production was achieved in the recombinant strain. Additionally, introduction and functional expression of the leech hyaluronidase in the recombinant strain substantially increased the yield of HA to 19.38 g·L−1. Moreover, HAs with a broad range of molecular weights (103 Da to 106 MDa) were efficiently produced by controlling the expression level of hyaluronidase using RBS mutants with different translational strengths. The method of the present invention can be used to produce low molecular weight HAs at large scale in industrial applications.

5. 发明申请

US20170175156A1 In vitro method for fast scarless DNA assembly using thermostable exonucleases and ligase 审中-公开
公开(公告)号：US20170175156A1
公开(公告)日：2017-06-22
申请号：US15347750
申请日：2016-11-09
申请人： Zhen Kang , Jian Chen , Peng Jin , Guocheng Du , Wenwen Ding
发明人： Zhen Kang , Jian Chen , Peng Jin , Guocheng Du , Wenwen Ding
IPC分类号： C12P19/34
CPC分类号： C12P19/34 , C12N15/1031 , C12Q1/6806 , C12Q2521/319 , C12Q2521/501 , C12Q2531/113
摘要： The present invention provides a method for scarless in vitro DNA assembly using thermostable exonucleases and ligase, which relates to the field of genetic engineering. The present invention provides a fast method for assembling DNA subfragments with homologous ends, which employs thermostable polymerases and ligase in a thermal cycle of denaturation, annealing, digestion and ligation. After denaturation, DNA subfragments are assembled together via annealing of the homologous end sequences, the unpaired single-stranded overhangs are digested by polymerases, and the resulting nicked gaps are sealed by a ligase. Using this method, 2-6 DNA subfragments were successfully assembled within two hours. This method can be used in conventional DNA recombination and be adapted to high throughput assembly operations. In addition, combinatorial mutations can be easily introduced into the assembled sequence by use of primers with mutated bases. It is particularly suitable for making enzyme and synthetic pathways mutation libraries with high diversity, which can be used in directed evolution to screen for enzymes and synthetic pathways with desirable properties.

6. 发明申请

US20170073719A1 Method of constructing a recombinant Bacillus subtilis that can produce specific-molecular-weight hyaluronic acids 有权
标题翻译：构建可产生特定分子量透明质酸的重组枯草芽孢杆菌的方法
公开(公告)号：US20170073719A1
公开(公告)日：2017-03-16
申请号：US15011509
申请日：2016-01-30
申请人： Jian Chen , Zhen Kang , Guocheng Du , Peng Jin
发明人： Jian Chen , Zhen Kang , Guocheng Du , Peng Jin
IPC分类号： C12P19/26 , C12N9/10 , C12N9/12 , C12N9/90 , C12N9/26 , C12N9/04
CPC分类号： C12P19/26 , C12N9/0006 , C12N9/1096 , C12N9/1241 , C12N9/2474 , C12N9/90 , C12Y101/01022 , C12Y206/01016 , C12Y207/07009 , C12Y207/07023 , C12Y302/01035 , C12Y504/0201
摘要： The present invention relates to the field of biotechnology engineering. It provides a method of constructing a recombinant Bacillus subtilis that can produce specific-molecular-weight hyaluronic acids. By integranted expression of hasA from Streptococcus zooepidemicus and overexpression of genes of HA synthetic pathway, tuaD, glmU and glmS, high yield HA production was achieved in the recombinant strain. Additionally, introduction and functional expression of the leech hyaluronidase in the recombinant strain substantially increased the yield of HA to 19.38 g·L−1. Moreover, HAs with a broad range of molecular weights (103 Da to 106 MDa) were efficiently produced by controlling the expression level of hyaluronidase using RBS mutants with different translational strengths. The method of the present invention can be used to produce low molecular weight HAs at large scale in industrial applications.
摘要翻译：本发明涉及生物技术工程领域。它提供了构建能产生特定分子量透明质酸的重组枯草芽孢杆菌的方法。通过来自猪链球菌的hasA的整合表达和HA合成途径，tuaD，glmU和glmS的基因的过表达，在重组菌株中实现了高产量的HA产生。此外，重组菌株中水蛭透明质酸酶的引入和功能表达显着提高了HA的产量至19.38g·L-1。此外，通过使用具有不同平移强度的RBS突变体控制透明质酸酶的表达水平，有效地产生具有宽范围分子量（103Da至106Mpa）的HA。本发明的方法可用于在工业应用中大规模生产低分子量的HAs。

7. 发明授权

US09279111B2 Leech hyaluronidase and its application 有权
标题翻译：水蛭透明质酸酶及其应用
公开(公告)号：US09279111B2
公开(公告)日：2016-03-08
申请号：US14251579
申请日：2014-04-12
申请人： Jian Chen , Guocheng Du , Zhen Kang , Peng Jin
发明人： Jian Chen , Guocheng Du , Zhen Kang , Peng Jin
IPC分类号： C12N9/24 , C12P19/26 , C12P21/02
CPC分类号： C12N9/2402 , C12P19/26 , C12P21/02 , C12Y302/01036
摘要： The present invention provides a novel leech HAase and a method of producing low-molecular-weight HA oligosaccharides using the leech HAase. This invention successfully cloned the first leech HAase gene and provides a method for high-level expression of the leech HAase gene. By controlling the incubation condition, different HA oligosaccharides, particularly HA4, HA6, HA8 and HA10, can be selectively generated using the leech HAase. The large-scale expression of the leech HAase and the enzymatic production of specific HA oligosaccharides are not only useful for the cosmetic, healthcare and the medical industries but also can be a great help to polysaccharides chemical synthesis and cancer research.
摘要翻译：本发明提供了一种新的水蛭HAase和使用该水蛭HAase生产低分子量HA寡糖的方法。本发明成功克隆了第一个水蛭HAase基因，提供了一种高水平表达水蛭HAase基因的方法。通过控制孵育条件，可以使用水蛭HAase选择性地产生不同的HA寡糖，特别是HA4，HA6，HA8和HA10。水蛭HAase的大规模表达和特异性HA寡糖的酶促生产不仅可用于化妆品，保健和医疗行业，而且可以有助于多糖化学合成和癌症研究。

8. 发明授权

US07283376B2 Mounting apparatus for expansion cards 失效
标题翻译：扩充卡安装装置
公开(公告)号：US07283376B2
公开(公告)日：2007-10-16
申请号：US11019329
申请日：2004-12-21
申请人： Shao-Bo Han , Zhen Kang
发明人： Shao-Bo Han , Zhen Kang
IPC分类号： H05K7/12
CPC分类号： H05K7/1418
摘要： A mounting apparatus for expansion cards (80) comprises a bracket (20), a pair of sliding members (40) and a retainer (60). The bracket staying in the chassis (90) of a computer comprises a main body (22), at the edge of the main body extends inwardly a first guiding part (224). A sliding portion (42) and a clasp portion (48) are formed on each sliding member. The sliding portion can engage in the first guiding part slidably, a post (482) protrudes outwardly from each clasp portion. The retainer comprises a rear wall (62), a serial of slots are defined on the rear wall successively for the posts to slide in. The retainer moves up and down when the posts slide in the slots.
摘要翻译：用于扩展卡（80）的安装装置包括托架（20），一对滑动构件（40）和保持器（60）。停留在计算机的底盘（90）中的支架包括主体（22），在主体的边缘处向内延伸有第一引导部分（224）。在每个滑动构件上形成滑动部分（42）和扣环部分（48）。滑动部分可滑动地接合在第一引导部分中，从每个扣环部分向外突出的柱子（482）。保持器包括后壁（62），连续地在后壁上限定了一些槽，用于柱滑动。当柱在槽中滑动时，保持器上下移动。

9. 发明申请

US20060092606A1 I/O interface cover assembly of computer enclosure 失效
标题翻译：计算机外壳的I / O接口盖组件
公开(公告)号：US20060092606A1
公开(公告)日：2006-05-04
申请号：US11267471
申请日：2005-11-04
申请人： Shao-Bo Han , Zhen Kang
发明人： Shao-Bo Han , Zhen Kang
IPC分类号： G06F1/16
CPC分类号： G06F1/16 , G06F1/181
摘要： An I/O interface cover assembly includes a panel, a bracket for accommodating an I/O interface element, a driving device, a locking device and a button. The panel defines an opening and a button hole adjacent the opening. The bracket is received in the opening, and includes one end pivotally connected to the panel and the other end defining an I/O slot and a locking hole therein. The driving device is disposed between the panel and the bracket for driving the bracket out from the opening of the panel. The locking device is movably arranged to the panel, and engagable with the locking hole of the bracket. The locking device is disposed with a resilient element. The button is disposed in the button hole of the panel, and is able to press the locking device to disengage from the locking hole of the bracket.
摘要翻译： I / O接口盖组件包括面板，用于容纳I / O接口元件的支架，驱动装置，锁定装置和按钮。面板限定开口和邻近开口的按钮孔。支架被容纳在开口中，并且包括一端枢转地连接到面板，另一端在其中限定I / O槽和锁定孔。驱动装置设置在面板和支架之间，用于从面板的开口驱动支架。锁定装置可移动地布置到面板上，并且可与支架的锁定孔接合。锁定装置设置有弹性元件。按钮设置在面板的按钮孔中，并且能够按压锁定装置以从支架的锁定孔脱离。

10. 发明授权

US07218514B2 I/O interface cover assembly of computer enclosure 失效
标题翻译：计算机外壳的I / O接口盖组件
公开(公告)号：US07218514B2
公开(公告)日：2007-05-15
申请号：US11267471
申请日：2005-11-04
申请人： Shao-Bo Han , Zhen Kang
发明人： Shao-Bo Han , Zhen Kang
IPC分类号： G06F1/16
CPC分类号： G06F1/16 , G06F1/181
摘要： An I/O interface cover assembly includes a panel, a bracket for accommodating an I/O interface element, a driving device, a locking device and a button. The panel defines an opening and a button hole adjacent the opening. The bracket is received in the opening, and includes one end pivotally connected to the panel and the other end defining an I/O slot and a locking hole therein. The driving device is disposed between the panel and the bracket for driving the bracket out from the opening of the panel. The locking device is movably arranged to the panel, and engagable with the locking hole of the bracket. The locking device is disposed with a resilient element. The button is disposed in the button hole of the panel, and is able to press the locking device to disengage from the locking hole of the bracket.
摘要翻译： I / O接口盖组件包括面板，用于容纳I / O接口元件的支架，驱动装置，锁定装置和按钮。面板限定开口和邻近开口的按钮孔。支架容纳在开口中，并且包括一端枢转地连接到面板，另一端在其中限定I / O槽和锁定孔。驱动装置设置在面板和支架之间，用于从面板的开口驱动支架。锁定装置可移动地布置到面板上，并且可与支架的锁定孔接合。锁定装置设置有弹性元件。按钮设置在面板的按钮孔中，并且能够按压锁定装置以从支架的锁定孔脱离。

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