OrangerV1, Main, Exploration, bibRecord, 000430

The codA gene as a negative selection marker in Citrus

Identifieur interne : 000430 ( Main/Exploration ); précédent : 000429; suivant : 000431

The codA gene as a negative selection marker in Citrus

Auteurs : Maria Luiza Peixoto De Oliveira [États-Unis] ; Ed Stover [États-Unis] ; James G. Thomson [États-Unis]

Source :

SpringerPlus [ 2193-1801 ] ; 2015.

RBID : PMC:4469690

Abstract

The use of positive selectable marker genes is widespread in plant genetic transformation allowing transgenic cells to grow while repressing non-transgenic cells. Negative selectable markers, on the contrary, allow the repression or ablation of transgenic cells. The codA gene of Escherichia coli encodes cytosine deaminase that hydrolyzes 5-fluorocytosine (5-FC) into the cytotoxic compound 5 fluorouracil. We tested the transgenic expression of the bacterial codA gene in citrus as a conditional negative selection marker, with the goal of selecting against plant tissues in which a transgenic cassette has not been successfully removed. We developed transgenic citrus lines containing the selection cassette, codA::nptII, driven by double enhanced CaMV35S promoter, verified by Southern blot analysis, RT-PCR, DsRed expression and subjected these transgenic lines to a 5-FC sensitivity assay. We found that, while non-transgenic citrus were unaffected by the presence of 5-FC, all of the transformed lines displayed symptoms of toxicity, indicating that the codA gene could be used as a negative selectable marker in Citrus, for post-transformation detection of the removal of undesired sequences.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4469690

DOI: 10.1186/s40064-015-1047-y
PubMed: 26090311
PubMed Central: 4469690

Affiliations:

Le document en format XML

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<front><div type="abstract" xml:lang="en"><p>The use of positive selectable marker genes is widespread in plant genetic transformation allowing transgenic cells to grow while repressing non-transgenic cells. Negative selectable markers, on the contrary, allow the repression or ablation of transgenic cells. The <italic>codA</italic>
 gene of <italic>Escherichia coli</italic>
 encodes cytosine deaminase that hydrolyzes 5-fluorocytosine (5-FC) into the cytotoxic compound 5 fluorouracil. We tested the transgenic expression of the bacterial <italic>codA</italic>
 gene in citrus as a conditional negative selection marker, with the goal of selecting against plant tissues in which a transgenic cassette has not been successfully removed. We developed transgenic citrus lines containing the selection cassette, <italic>codA</italic>
::<italic>nptII</italic>
, driven by double enhanced <italic>CaMV35S</italic>
 promoter, verified by Southern blot analysis, RT-PCR, <italic>DsRed</italic>
 expression and subjected these transgenic lines to a 5-FC sensitivity assay. We found that, while non-transgenic citrus were unaffected by the presence of 5-FC, all of the transformed lines displayed symptoms of toxicity, indicating that the <italic>codA</italic>
 gene could be used as a negative selectable marker in Citrus, for post-transformation detection of the removal of undesired sequences.</p>
</div>
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The codA gene as a negative selection marker in Citrus

The codA gene as a negative selection marker in Citrus

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