• 제목/요약/키워드: dapD gene

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Brevibacterium lactofermentum의 dapD 유전자의 Cloning 및 E. coli에서의 발현 (Cloning and Expression of the dapD Gene from Brevibacterium lactofermentum in E. coli)

  • 김옥미;박선희;박혜경;이승언;하대중;이갑랑
    • 한국식품영양과학회지
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    • 제30권5호
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    • pp.802-805
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    • 2001
  • 산업적으로 lysine 발효 산업에 이용되고 있는 B. lactofermentum으로부터 lysine 생합성에 관여하는 tetrahyrodipicolinate N-succinyl transferase를 지령하는 dapD 유전자를 E. coli의 dapD 결손변이주와의 complementation test를 통하여 cloning하였다. 재조합 plamid는 3.6 kb의 DNA 단편을 함유하고 있었으며 Southern blot hybridization을 통하여 dapD 유전자는 B. lactofermentum으로부터 유래하였으며 염색체 DNA내에 single copy로 존재함을 알 수 있었다. 또한 lysine 생성량 분석을 통하여 E. coli에서 B. lactofermentum dapD 유전자의 발현을 확인하였다.

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Cloning and Sequencing of the ddh Gene involved in the Novel Pathway of Lysine Biosynthesis from Brevibacterium lactofermentum

  • Kim, Ok-Mi;Kim, Hyun-Jeong;Kim, Dal-Sang;Park, Dong-Chul;Lee, Kap-Rang
    • Journal of Microbiology and Biotechnology
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    • 제5권5호
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    • pp.250-256
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    • 1995
  • The ddh gene encoding meso-diaminopimelate (meso-DAP)-dehydrogenase (DDH) in Brevibacterium lactofermentum was isolated by complementation of the Escherichia coli dapD mutation. It was supposed from subcloning experiments and complementation tests that the evidence for DDH activity appeared in about 2.5 kb Xhol fragmented genome. The 2.5 kb Xhol fragment containing the ddh gene was sequenced, and an open reading frame of 960 bp encoding a polypeptide comprising 320 amino acids was found. Computer analysis indicated that the deduced amino acid of the B. lactofermentum ddh gene showed a high homology with that of the Corynebacterium glutamicum ddh gene.

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Development of a Highly Active Fluorescence-Based Detector for Yeast G Protein-Coupled Receptor Ste2p

  • Hong, Jin Woo;Ahn, Hee Jun;Baek, Jee Su;Hong, Eun young;Jin, Dong Hoon;Khang, Yong Ho;Hong, Nam Joo
    • Journal of Microbiology and Biotechnology
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    • 제28권10호
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    • pp.1589-1603
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    • 2018
  • Twenty analogs of $[Orn^6,D-Ala^9]{\alpha}-factor$ were synthesized and assayed for their biological activities: seven analogs of $[Orn^6,X^9]{\alpha}-factor$, seven analogs of $[X^6,D-Ala^9]{\alpha}-factor$, five analogs of $[X^5,X^6,D-Ala^9]{\alpha}-factor$, and native ${\alpha}-factor$ (X = amino acids). Their biological activities (halo, gene induction, and affinity) were measured using S. cerevisiae Y7925 and LM102 and compared with those of native ${\alpha}-factor$ (100%). G protein-coupled receptor was expressed in strain LM102 containing pESC-LEU-STE2 vector. $[Dap^6,D-Ala^9]{\alpha}-factor$ with weak halo activity (10%) showed the highest receptor affinity (> 230%) and the highest gene induction activity (167%). $[Arg^6,D-Ala^9]{\alpha}-factor$ showed the highest halo activity (2,000%). The number of active binding sites per cell (about 20,000 for strain LM102) was determined using a newly-designed fluorescence-based detector, $[Arg^6,D-Ala^9]{\alpha}-factor-Edan$, with high sensitivity (12,500-fold higher than the absorption-based detector $[Orn^6]{\alpha}-factor-[Cys]_3$).