Expression of a synthetic cholera toxin B subunit in tobacco using ubiquitin promoter and bar gene as a selectable marker.

Abstract:

:A protocol has been developed to produce a cholera toxin B subunit (CTB) in tobacco tolerant to the herbicide phosphinothricin (PPT) by means of in vitro selection. The synthetic CTB subunit gene was altered to modify the codon usage to that of tobacco plant genes. The gene was then cloned into a plant expression vector and was under the control of the ubiquitin promoter and transformed into tobacco plants by Agrobacterium-mediated transformation. Transgenic plantlets were selected in a medium supplemented with 5 mg/L PPT. Polymerase chain reaction analysis confirmed stable integration of the synthetic CTB gene into a chromosomal DNA. A high level of CTB (1.8% of total soluble protein) was expressed in transgenic plants, which was 18-fold higher than that under the control of the expressed CaMV 35S promoter with native gene. The transgenic plants when transferred to a greenhouse proved to be resistant to 2% PPT.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Kang TJ,Kim BG,Yang JY,Yang MS

doi

10.1385/MB:32:2:093

keywords:

subject

Has Abstract

pub_date

2006-02-01 00:00:00

pages

93-100

issue

2

eissn

1073-6085

issn

1559-0305

pii

MB:32:2:093

journal_volume

32

pub_type

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