4-Methylumbelliferyl beta-D-glucuronide → 4-Methylumbelliferone

HRID 458504

Reaction equation

HRID 458504 reaction equation
[CH3:1][C:2]1[C:8]2[CH:9]=[CH:10][C:11]([O:13][C@@H]3O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]3O)=[CH:12][C:7]=2[O:6][C:4](=[O:5])[CH:3]=1>C([O-])([O-])=O.[Na+].[Na+]>[CH3:1][C:2]1[C:8]2[CH:9]=[CH:10][C:11]([OH:13])=[CH:12][C:7]=2[O:6][C:4](=[O:5])[CH:3]=1

Participants

3

Reactant 1

HCID 915534-Methylumbelliferyl beta-D-glucuronide

Product 1

HCID 52805674-Methylumbelliferone

Reagents, catalysts & solvents 1

HCID 10340Sodium CarbonateSolvent

Procedure

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 7.01, 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 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 BIORAD Protein Assay kit, with serum albumin as the standard. Three seeds were assayed for each transgenic plant.

Workup

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

Sources & evidence

ORD record
ord-2531acde295a4a6a89b31f4c70c2fcb2
Source dataset
uspto-grants-2007_03
Patent number
US07192774B2

Linked notes

Loading…