公开(公告)号：US5219738A

公开(公告)日：1993-06-15

申请号：US45512

申请日：1987-05-04

申请人： Noboru Tomioka ,  Fumio Omae ,  Eriko Mine ,  Tomoko Ishii

发明人： Noboru Tomioka ,  Fumio Omae ,  Eriko Mine ,  Tomoko Ishii

IPC分类号： C12N9/50 ,  A61K38/00 ,  C12N5/00 ,  C12N5/10 ,  C12N9/64 ,  C12N9/72 ,  C12N15/00 ,  C12N15/09 ,  C12N15/85 ,  C12P21/00 ,  C12R1/91

CPC分类号： C12N9/6459 ,  C12N15/85 ,  C12Y304/21069 ,  A61K38/00

摘要： By screening an animal cell transformant, which has been obtained by introducing into wild-type animal cells a wild-type DHFR gene and a structural gene which codes a desired useful substance, with a medium containing methotrexate (MTX) at a concentration of 50 .mu.M or lower, both genes can be amplified effectively within an MTX resistant strain and a stable amplified transformant can also be obtained. The screening may be repeated using an increasing concentration of MTX. It is also possible to obtain stably the desired foreign gene product in a high yield by using the thus-obtained amplified transformant.

公开(公告)号：US4824782A

公开(公告)日：1989-04-25

申请号：US180691

申请日：1988-04-08

申请人： Yoshio Furutani ,  Noboru Tomioka ,  Masaru Honjo ,  Kazuaki Manabe ,  Hiroaki Shimada

发明人： Yoshio Furutani ,  Noboru Tomioka ,  Masaru Honjo ,  Kazuaki Manabe ,  Hiroaki Shimada

IPC分类号： C12N15/09 ,  C07H21/04 ,  C12N1/20 ,  C12N1/21 ,  C12N9/56 ,  C12N15/00 ,  C12N15/67 ,  C12N15/75 ,  C12P21/00 ,  C12R1/07 ,  C12N7/00

CPC分类号： C12N9/54 ,  C12N15/67 ,  C12N15/75

摘要： This invention relates to a DNA base sequence, capable of increasing the amount of protein secreted by microorganisms, and its derivative sequences; a recombinant plasmid including the whole or a part of said DNA base sequence; a method for preparing the recombinant plasmid in which, when microorganisms having introduced thereinto a recombinant DNA including the desired DNA base sequence are to be separated in the process of cloning, suitable transformants can be efficiently selected by taking the amount of protein secreted out of the cells and particularly the activity of an enzyme protein as an index; and a method of microbial breeding which comprises introducing the recombinant plasmid into a microorganism to increase the amount of protein secreted by the microorganism.

摘要翻译： 本发明涉及能够增加微生物分泌蛋白质的DNA碱基序列及其衍生序列; 包含全部或部分所述DNA碱基序列的重组质粒; 一种重组质粒的制备方法，其中，在将包含所需DNA碱基序列的重组DNA导入其中的微生物在克隆过程中被分离的情况下，可以通过从所述重组质粒中分泌出的蛋白质的量来有效地选择合适的转化体 细胞，特别是酶蛋白的活性作为指标; 以及微生物育种方法，其包括将重组质粒导入微生物以增加微生物分泌的蛋白质的量。

公开(公告)号：US07752917B2

公开(公告)日：2010-07-13

申请号：US10556388

申请日：2003-05-12

申请人： Noboru Tomioka

发明人： Noboru Tomioka

IPC分类号： G01N3/24

CPC分类号： G06F17/5018 ,  G01N3/00 ,  G01N2203/0073 ,  G01N2203/0214 ,  G01N2203/0216 ,  G06F2217/42 ,  G06F2217/76

摘要： A fatigue life estimating method for a spot welded structure is provided comprising the steps of providing a shell model of a spot welded structure for a finite element method analyzing process, calculating the nominal structural stress on a nugget as the center of the spot welded structure using a disk bending theory and a two-dimensional elastic theory of the elastodynamics with the partial loads exerted on the nugget and the deflection on the circumference of a circle, D in diameter, in which the nugget is located, which have been calculated by the finite element method analyzing process of the shell model, and estimating the fatigue life of the spot welded structure from the nominal structure stress. According to the method, the fatigue life of the spot welded structure can be estimated easily, readily, and accurately.

摘要翻译： 提供一种点焊结构的疲劳寿命估计方法，包括以下步骤：提供用于有限元法分析过程的点焊结构的壳模型，计算熔核上的标称结构应力作为点焊结构的中心，使用 圆盘弯曲理论和弹性动力学的二维弹性理论，其中施加在熔核上的部分载荷和圆形圆周上的弯曲，直径（D），其中熔核位于其中，其由有限的 模型的元素分析过程，以及从标称结构应力估计点焊结构的疲劳寿命。 根据该方法，可以容易且准确地估计点焊结构的疲劳寿命。

公开(公告)号：US20090211366A1

公开(公告)日：2009-08-27

申请号：US10556388

申请日：2003-05-12

申请人： Noboru Tomioka

发明人： Noboru Tomioka

IPC分类号： G01N3/24

CPC分类号： G06F17/5018 ,  G01N3/00 ,  G01N2203/0073 ,  G01N2203/0214 ,  G01N2203/0216 ,  G06F2217/42 ,  G06F2217/76

摘要： A fatigue life estimating method for a spot welded structure is provided comprising the steps of providing a shell model of a spot welded structure for a finite element method analyzing process, calculating the nominal structural stress on a nugget as the center of the spot welded structure using a disk bending theory and a two-dimensional elastic theory of the elastodynamics with the partial loads exerted on the nugget and the deflection on the circumference of a circle, D in diameter, in which the nugget is located, which have been calculated by the finite element method analyzing process of the shell model, and estimating the fatigue life of the spot welded structure from the nominal structure stress.According to the method, the fatigue life of the spot welded structure can be estimated easily, readily, and accurately.

摘要翻译： 提供一种点焊结构的疲劳寿命估计方法，包括以下步骤：提供用于有限元法分析过程的点焊结构的壳模型，计算熔核上的标称结构应力作为点焊结构的中心，使用 圆盘弯曲理论和弹性动力学的二维弹性理论，其中施加在熔核上的部分载荷和圆形圆周上的弯曲，直径（D），其中熔核位于其中，其由有限的 模型的元素分析过程，以及从标称结构应力估计点焊结构的疲劳寿命。 根据该方法，可以容易且准确地估计点焊结构的疲劳寿命。

公开(公告)号：US4731327A

公开(公告)日：1988-03-15

申请号：US896583

申请日：1986-08-18

申请人： Yoshio Furutani ,  Masaru Honjo ,  Kazuaki Manabe ,  Hiroaki Shimada ,  Noboru Tomioka

发明人： Yoshio Furutani ,  Masaru Honjo ,  Kazuaki Manabe ,  Hiroaki Shimada ,  Noboru Tomioka

IPC分类号： C12N15/09 ,  C12N9/54 ,  C12N15/67 ,  C12N15/75 ,  C12N15/00 ,  C12P21/00

CPC分类号： C12N15/75 ,  C12N15/67 ,  C12N9/54

摘要： A recombinant DNA molecule, a portion of which includes a vector whose replication is initiated independently of the temperature-sensitive mutant gene of a temperature-sensitive mutant strain of the genus Bacillus as host is introduced into the host. The host is then cultured at a temperature which does not completely inhibit chromosomal DNA replication. By this process, the recombinant DNA molecule can be stabilized in the host and its expression can be enhanced.

摘要翻译： 重组DNA分子，其一部分包括其复制独立于作为宿主的芽孢杆菌属的温度敏感突变菌株的温度敏感性突变基因启动的载体导入宿主。 然后在不完全抑制染色体DNA复制的温度下培养宿主。 通过该方法，重组DNA分子可以在宿主中稳定，并且其表达可以增强。

公开(公告)号：US5612192A

公开(公告)日：1997-03-18

申请号：US359863

申请日：1994-12-20

申请人： Yoshio Furutani ,  Hiroaki Shimada ,  Izumi Mita ,  Kazuaki Manabe ,  Masaru Honjo ,  Noboru Tomioka

发明人： Yoshio Furutani ,  Hiroaki Shimada ,  Izumi Mita ,  Kazuaki Manabe ,  Masaru Honjo ,  Noboru Tomioka

IPC分类号： C12N9/54 ,  C12N15/75 ,  C12N15/00 ,  C12P21/00

CPC分类号： C12N15/75 ,  C12N9/54 ,  C07K2319/02 ,  C07K2319/036

摘要： A neutral protease gene of Bacillus amyloliquefaciens is cloned, which gene comprises the promoter region, the ribosome binding region, the region involved in the secretion of the neutral protease, the region consisting of the structural gene for the neutral protease, and the terminator region. Each of the regions is useful as a material for construction of a recombinant DNA used for the production of proteins by culturing a host bacterium transformed with the recombinant DNA. For example, the extracellular production of neutral protease in a large amount can be accomplished by culturing B. subtilis transformed with a recombinant DNA comprising pUB110 and the neutral protease gene.

摘要翻译： 克隆解淀粉芽孢杆菌的中性蛋白酶基因，该基因包含启动子区，核糖体结合区，参与中性蛋白酶分泌的区域，由中性蛋白酶的结构基因组成的区域和终止区。 通过培养用重组DNA转化的宿主细菌，每个区域可用作构建用于生产蛋白质的重组DNA的材料。 例如，可以通过培养用包含pUB110和中性蛋白酶基因的重组DNA转化的枯草芽孢杆菌来实现大量的中性蛋白酶的细胞外产生。