The effect of Ca-formaldehyde fixation on the activity of the reporter enzyme β-glucuronidase (GUS) in transgenic tobacco

The inhibition of β-glucuronidase (GUS) activity by Ca-formaldehyde fixation was investigated in leaves of transgenic tobacco ( Nicotiana tabacum L.) expressing the β-glucuronidase uidA reporter gene from Escherichia coli. One-hour fixation of a freefloating leaf disc in a cold fixative decreased th...

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Bibliographic Details
Published in:Acta histochemica 1995-04, Vol.97 (2), p.137-140
Main Authors: Vitha, Stanislav, Beneš, Karel, Michalová-Voldánová, Marta
Format: Article
Language:English
Subjects:
Calcium
Escherichia coli - enzymology
Escherichia coli - genetics
fixation
Formaldehyde
Genes, Bacterial
Glucuronidase - analysis
Glucuronidase - biosynthesis
Histological Techniques
Indicators and Reagents
Nicotiana - cytology
Nicotiana - enzymology
Nicotiana tabacum
Plants, Genetically Modified
Plants, Toxic
transgenosis
β-glucuronidase
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Summary:The inhibition of β-glucuronidase (GUS) activity by Ca-formaldehyde fixation was investigated in leaves of transgenic tobacco ( Nicotiana tabacum L.) expressing the β-glucuronidase uidA reporter gene from Escherichia coli. One-hour fixation of a freefloating leaf disc in a cold fixative decreased the activity of GUS by 60070 as compared with the untreated control disc kept in a moist chamber. The inhibition was identical in the periphery and centre of the disc. Fixation including initial vacuum infiltration of the fixative led to almost complete inactivation of the enzyme. The incubation of discs in H 2O, either with or without vacuum infiltration, did not lower GUS activity.
ISSN:0065-1281
1618-0372
DOI:10.1016/S0065-1281(11)80089-9