公开(公告)号：EP1219712B1

公开(公告)日：2008-05-21

申请号：EP00964654.8

申请日：2000-10-04

申请人： Ajinomoto Co., Inc.

发明人： HIRANO,Seiko Ajinomoto Co.Inc. Fermentat.&Bio.Lab. ,  NONAKA,Gen Ajinomoto Co.Inc. Fermentation&Bio.Lab. ,  MATSUZAKI,Yumi Ajinomoto Co.Inc. Fermenta.&Bio.Lab ,  AKIYOSHI,Naoki Ajinomoto Co.Inc. Fermenta.&Bio.Lab ,  NAKAMURA,Kanae 902 Stamford Court 8A ,  KIMURA,Eiichiro Ajinomoto Co.Inc. Ferment.&Bio.Lab ,  OSUMI,Tsuyoshi Ajinomoto Co.Inc. Fermenta.&Bio.Lab ,  MATSUI,Kazuhiko Ajinomoto Co.Inc. Fermenta.Bio.Lab ,  KAWAHARA,Yoshio Ajinomoto Co.Inc. Fermenta.Bio.Lab ,  KURAHASHI,Osamu Ajinomoto Co.Inc. Fermenta.Bio.Lab ,  NAKAMATSU,Tsuyoshi Ajinomoto Co.Inc. Ferme.Bio.Lab ,  SUGIMOTO,Shinichi Ajinomoto Co.Inc. Fermen.Bio.Lab

IPC分类号： C12N15/60 ,  C12N15/54 ,  C12N15/53 ,  C12N15/31 ,  C12N15/56 ,  C12N9/88 ,  C12N9/12 ,  C12N9/04 ,  C07K14/34 ,  C12N9/26 ,  C12P13/04

CPC分类号： C12Y108/01004 ,  C07K14/34 ,  C12N9/0006 ,  C12N9/0008 ,  C12N9/0051 ,  C12N9/1205 ,  C12N9/2408 ,  C12N9/88 ,  C12Y101/01041 ,  C12Y102/01051 ,  C12Y102/04002 ,  C12Y104/01004 ,  C12Y203/03001 ,  C12Y207/01011 ,  C12Y302/01026 ,  C12Y401/01031 ,  C12Y401/03001 ,  C12Y402/01003 ,  C12Y604/01001 ,  C12Y604/01002

公开(公告)号：EP1219712A1

公开(公告)日：2002-07-03

申请号：EP00964654.8

申请日：2000-10-04

申请人： Ajinomoto Co., Inc.

发明人： HIRANO,Seiko Ajinomoto Co.Inc. Fermentat.&Bio.Lab. ,  NONAKA,Gen Ajinomoto Co.Inc. Fermentation&Bio.Lab. ,  MATSUZAKI,Yumi Ajinomoto Co.Inc. Fermenta.&Bio.Lab ,  AKIYOSHI,Naoki Ajinomoto Co.Inc. Fermenta.&Bio.Lab ,  NAKAMURA,Kanae 902 Stamford Court 8A ,  KIMURA,Eiichiro Ajinomoto Co.Inc. Ferment.&Bio.Lab ,  OSUMI,Tsuyoshi Ajinomoto Co.Inc. Fermenta.&Bio.Lab ,  MATSUI,Kazuhiko Ajinomoto Co.Inc. Fermenta.Bio.Lab ,  KAWAHARA,Yoshio Ajinomoto Co.Inc. Fermenta.Bio.Lab ,  KURAHASHI,Osamu Ajinomoto Co.Inc. Fermenta.Bio.Lab ,  NAKAMATSU,Tsuyoshi Ajinomoto Co.Inc. Ferme.Bio.Lab ,  SUGIMOTO,Shinichi Ajinomoto Co.Inc. Fermen.Bio.Lab

IPC分类号： C12N15/60 ,  C12N15/54 ,  C12N15/53 ,  C12N15/31 ,  C12N15/56 ,  C12N9/88 ,  C12N9/12 ,  C12N9/04 ,  C07K14/34 ,  C12N9/26 ,  C12P13/04

CPC分类号： C12Y108/01004 ,  C07K14/34 ,  C12N9/0006 ,  C12N9/0008 ,  C12N9/0051 ,  C12N9/1205 ,  C12N9/2408 ,  C12N9/88 ,  C12Y101/01041 ,  C12Y102/01051 ,  C12Y102/04002 ,  C12Y104/01004 ,  C12Y203/03001 ,  C12Y207/01011 ,  C12Y302/01026 ,  C12Y401/01031 ,  C12Y401/03001 ,  C12Y402/01003 ,  C12Y604/01001 ,  C12Y604/01002

摘要： A plurality of primer sets are designed based on a region where conservation at the amino acid level is observed among various microorganisms for known gene sequences corresponding to a gene coding for an enzyme of the L-amino acid biosynthetic pathway derived from Corynebacterium thermoaminogenes, preferably an enzyme that functions at a higher temperature compared with that of Corynebacterium glutamicum. PCR is performed by using the primers and chromosomal DNA of Corynebacterium thermoaminogenes as a template. The primers with which an amplification fragment has been obtained are used as primers for screening to select a clone containing a target DNA fragment from a plasmid library of chromosomal DNA of Corynebacterium thermoaminogenes.

摘要翻译： 基于对应于编码来源于热棒氨酸棒杆菌的L-氨基酸生物合成途径的酶的基因的已知基因序列的各种微生物在各种微生物中观察到氨基酸水平保护的区域来设计多个引物组，优选为 与谷氨酸棒杆菌相比在更高的温度下起作用的酶。 通过使用热源氨基酸棒杆菌的引物和染色体DNA作为模板进行PCR。 将获得了扩增片段的引物用作筛选引物，从嗜热氨基酸棒杆菌的染色体DNA的质粒文库中选择含有靶DNA片段的克隆。