反应 #1702043

ord-f92868cf746c477e95e4d51955ed4812

反应方程式

CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
IPTG
CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
IPTG
C[C@H](Nc1ccc(O)c(O)c1)C(=O)O
DOPA
C[C@H](Nc1ccc(O)c(O)c1)C(=O)O
DOPA
CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
IPTG
N[C@@H](Cc1ccc(O)cc1)C(=O)O
tyrosine
C[C@H](Nc1ccc(O)c(O)c1)C(=O)O
DOPA
→
N[C@@H](Cc1ccc(O)c(O)c1)C(=O)O
DOPA

反应物

CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
IPTG
CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
IPTG
C[C@H](Nc1ccc(O)c(O)c1)C(=O)O
DOPA
C[C@H](Nc1ccc(O)c(O)c1)C(=O)O
DOPA
CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
IPTG
N[C@@H](Cc1ccc(O)cc1)C(=O)O
tyrosine
C[C@H](Nc1ccc(O)c(O)c1)C(=O)O
DOPA

试剂

无

反应条件

详细条件
See reaction.notes.procedure_details.

后处理

  1. 1
    workup.ADDITIONwas introduced to the concentration of 1 mM in the medium
  2. 2
    workup.ADDITIONwas added
  3. 3
    workup.ADDITIONfor a negative control, nothing was introduced
  4. 4
    workup.WAITAfter culture, the cells were centrifuged at 4000 rpm for 10 minutes
  5. 5
    其他supernatant was removed
  6. 6
    其他the cells were recovered

实验过程

After confirming stationary phase, DOPA (3,4-dihydroxyphenyl-L-alanine) was introduced to the concentration of 1 mM in the medium. After introducing DOPA, inducer IPTG(isopropyl-β-D-thiogalactopyranoside, 1 mM) was added to induce the expression of the above proteins. After adding IPTG, culture was conducted at 37° C. for additional 6 hours. For a positive control, tyrosine was introduced to the concentration of 1 mM instead of DOPA, and the expression of the proteins was induced with IPTG, and for a negative control, nothing was introduced. After culture, the cells were centrifuged at 4000 rpm for 10 minutes, and then, supernatant was removed, and the cells were recovered.

来源

DOI: 10.6084/m9.figshare.5104873.v1专利: US08765682B2uspto-grants-2014_07