β-glucuronidase gene expression and mRNA stability in oat protoplasts.

Abstract:

:Protoplasts derived from oat (Avena sativa L.) suspension culture cells (7 days after subculturing) were electroporated with plasmid DNA containing the Escherichia coli uidA gene encoding the ß-glucuronidase reporter enzyme. Consistently high enzyme activity was observed with electroporation conditions of 500 μF and 1125 volts/cm. Enzyme activity and mRNA accumulation time courses were determined. The maximum enzyme activity was detected at 24 hours after electroporation, while the maximum mRNA level was detected at 12 hours after electroporation. ß-glucuronidase mRNA was in vitro synthesized with and without a 5' methylated cap and then electroporated into protoplasts. Only capped mRNA produced significant enzyme activity. By electroporating radiolabeled, in vitro synthesized mRNA, the ß-glucuronidase mRNA half-life was estimated to be ∼35 minutes in oat protoplasts.

journal_name

Plant Cell Rep

journal_title

Plant cell reports

authors

Higgs DC,Colbert JT

doi

10.1007/BF00234710

subject

Has Abstract

pub_date

1993-05-01 00:00:00

pages

445-52

issue

7-8

eissn

0721-7714

issn

1432-203X

journal_volume

12

pub_type

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