JP2003061687A5 - - Google Patents
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- JP2003061687A5 JP2003061687A5 JP2002151981A JP2002151981A JP2003061687A5 JP 2003061687 A5 JP2003061687 A5 JP 2003061687A5 JP 2002151981 A JP2002151981 A JP 2002151981A JP 2002151981 A JP2002151981 A JP 2002151981A JP 2003061687 A5 JP2003061687 A5 JP 2003061687A5
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- dapa
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RSFD80に含まれている変異型dapAは野生型dapAの塩基配列において塩基番号623のＣがＴに変化した配列を有し、それによって、コードされる変異型DDPSは118位のヒスチジン残基がチロシン残基に置換された配列を有する。また、RSFD80に含まれている変異型lysCは野生型lysCの塩基配列において塩基番号1638のＣがＴ変化した配列を有し、それによって、コードされる変異型AKIIIは352位のスレオニン残基がイソロイシン残基に置換された配列を有する。 Mutant dapA contained in RSFD80 has a sequence C at the nucleotide number 623 is changed to T in the base sequence of the wild type dapA, whereby the histidine residues of mutant DDPS is 1 # 18 encoded Has a sequence substituted with a tyrosine residue. Moreover, mutant lysC contained in RSFD80 has a sequence C is T changes at nucleotides 1638 in the nucleotide sequence of the wild type lysC, thereby encoded mutant AKIII is 3 52-position threonine residue It has a sequence in which the group is substituted with an isoleucine residue.
dapA*遺伝子断片は、同遺伝子を含む公知のプラスミドRSFD80(WO 95/16042号参照)を鋳型として、配列番号３および４に示すプライマーを用いたPCR法（変性94℃-20秒、アニーリング55℃-30秒、伸長反応72℃-60秒）により増幅した。PCR反応には、Pyrobest DNA polymerase（宝酒造社製）を使用した。得られたdapA*断片をPCRprep（Promega社製）にて精製した後、制限酵素Sse8387IおよびXbaIで消化した。フェノール・クロロホルム溶液を加えて混合し、反応を停止させた。反応液を遠心分離した後、上層を回収し、エタノール沈殿にてDNAを回収し、0.8%アガロースゲルで分離後、約0.1kbpのDNA断片を回収した。 dapA * gene fragment, as known plasmid RSFD80 (WO 95/16042 No. see) the mold containing the gene, SEQ ID NO: 3 and PCR using the primers shown in 4 (denaturation 94 ° C. -20 seconds, annealing 55 ° C. It was amplified by -30 seconds, extension reaction 72 ° C-60 seconds). Pyrobest DNA polymerase (manufactured by Takara Shuzo Co., Ltd.) was used for the PCR reaction. The obtained dapA * fragment was purified by PCRprep (manufactured by Promega) and then digested with restriction enzymes Sse8387I and XbaI. Phenol / chloroform solution was added and mixed, and the reaction was stopped. After centrifuging the reaction solution, the upper layer was recovered, DNA was recovered by ethanol precipitation, separated by 0.8% agarose gel, and then a DNA fragment of about 0.1 kbp was recovered.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2002151981A JP4120269B2 (en) | 2001-06-12 | 2002-05-27 | Method for producing L-lysine or L-arginine using methanol-assimilating bacteria |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2001-177075 | 2001-06-12 | ||
| JP2001177075 | 2001-06-12 | ||
| JP2002151981A JP4120269B2 (en) | 2001-06-12 | 2002-05-27 | Method for producing L-lysine or L-arginine using methanol-assimilating bacteria |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2003061687A JP2003061687A (en) | 2003-03-04 |
| JP2003061687A5 true JP2003061687A5 (en) | 2005-08-18 |
| JP4120269B2 JP4120269B2 (en) | 2008-07-16 |
Family
ID=26616761
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2002151981A Expired - Fee Related JP4120269B2 (en) | 2001-06-12 | 2002-05-27 | Method for producing L-lysine or L-arginine using methanol-assimilating bacteria |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP4120269B2 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4836440B2 (en) * | 2004-03-10 | 2011-12-14 | センター・フォー・ディーエヌエイ・フィンガープリンティング・アンド・ダイアグノスティックス | Method for producing arginine using microorganisms |
| MX2007014175A (en) | 2005-05-12 | 2008-01-14 | Ajinomoto Kk | Method for producing protein. |
| JP2009089603A (en) * | 2006-02-02 | 2009-04-30 | Ajinomoto Co Inc | Method for producing l-lysine using methanol-assimilating bacterium |
| JP2009153382A (en) | 2006-03-30 | 2009-07-16 | Ajinomoto Co Inc | Method for producing carboxylic acid using methanol-assimilating bacterium |
| KR102139806B1 (en) * | 2020-02-13 | 2020-07-30 | 씨제이제일제당 (주) | Microorganism Comprising Mutated LysE and Method of L-Amino Acid Production Using the Same |
-
2002
- 2002-05-27 JP JP2002151981A patent/JP4120269B2/en not_active Expired - Fee Related
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