2-Mercaptoethanol → 4-Methylumbelliferone

HRID 1436968

Reaktionsgleichung

Reaktionsgleichung von HRID 1436968
SCCO.C(N(CC([O-])=O)CC(O)=O)CN(CC([O-])=O)CC(O)=O.[Na+].[Na+].CCCCCCCCCCCCOS([O-])(=O)=O.[Na+].CCC(COC(C(N(CC[NH+](C)C)C)=O)(C1C=CC=CC=1)C1C=CC=CC=1)CC.[Cl-].[CH3:75][C:76]1[C:82]2[CH:83]=[CH:84][C:85]([O:87][C@@H]3O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]3O)=[CH:86][C:81]=2[O:80][C:78](=[O:79])[CH:77]=1>C([O-])([O-])=O.[Na+].[Na+]>[CH3:75][C:76]1[C:82]2[CH:83]=[CH:84][C:85]([OH:87])=[CH:86][C:81]=2[O:80][C:78](=[O:79])[CH:77]=1

Komponenten

7

Versuchsvorschrift

To evaluate the activity of various promoters, GUS fluorescence was assayed by the method of Jefferson (1987). Maturing seeds at 17 DAF were homogenized in GUS extraction buffer (50 mM NaPO4 [pH 7.0], 10 mM 2-mercaptoethanol, 10 mM Na2-EDTA, 0.1% SDS, 0.1% Triton X-100). After centrifugation, 10 μl of the supernatant was mixed with 90 μl of assay buffer containing 1 mM 4-methylumbelliferyl-β-D-glucuronide (MUG). After incubating for one hour at 37° C., 900 μl of 0.2 M Na2CO3 was added to the mixture to terminate the reaction. Values obtained using a fluorometer were compared with those obtained from serial dilutions of 4-methylumbelliferone (4MU). The protein amount was determined using a BIO-RAD® Protein Assay kit, which is based on the Bradford method (Bradford, M., Anal. Biochem. 72, 248 (1976)), with serum albumin as the standard. Three seeds were assayed for each transgenic plant.

Aufarbeitung

  1. additionAfter centrifugation, 10 μl of the supernatant was mixed with 90 μl of assay
  2. customto terminate
  3. customthe reaction
  4. customValues obtained

Quellen und Nachweise

ORD-Eintrag
ord-fe140407f7ab4885918f418d47bdaca3
Quelldatensatz
uspto-grants-2009_11
Patentnummer
US07619135B2

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