Articles
AGROBACTERIUM-MEDIATED TRANSFORMATION OF KALANCHOE BLOSSFELDIANA AND QUANTITATIVE ANALYSIS OF THE EXPRESSION PATTERN OF β-GLUCURONIDASE GENE
Article number
454_31
Pages
273 – 280
Language
Abstract
Leaf segments were infected with the Agrobacterium strain LBA4404 having a binary vector plasmid pBI121 or its derivatives and cultured on Murashige and Skoog medium supplemented with 0.5 mg/L benzyladenine, 2 mg/L indoleacetic acid, 300 mg/L carbenicillin and 20 mg/L kanamycin (selection medium) for regeneration.
Primary transformants with three kinds of chimeric
-glucuronidase (GUS) genes were obtained in different experiments, and Southern blot analysis confirmed the presence of the GUS gene in their genome.
The integrated GUS gene(s) seemed to be silenced in about half of the transformants.
There was no clear correlation between copy number and expression level of the GUS gene.
Observed levels of GUS activity within transformants were divided into three classes: 1) high activity, 2) low activity and 3) near-zero activity.
A stable method for Agrobacterium– mediated transformation of Kalanchoe blossfeldiana Poelln. (Crassulaceae) is reported.
Primary transformants with three kinds of chimeric
-glucuronidase (GUS) genes were obtained in different experiments, and Southern blot analysis confirmed the presence of the GUS gene in their genome.The integrated GUS gene(s) seemed to be silenced in about half of the transformants.
There was no clear correlation between copy number and expression level of the GUS gene.
Observed levels of GUS activity within transformants were divided into three classes: 1) high activity, 2) low activity and 3) near-zero activity.
A stable method for Agrobacterium– mediated transformation of Kalanchoe blossfeldiana Poelln. (Crassulaceae) is reported.
Authors
R. Aida, M. Shibata
Keywords
transgene silencing, copy number, expression pattern
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