Developmentally regulated site-specific marker gene excision in transgenic B. napus plants

Autor: Kopertekh, L.; Broer, I.; Schiemann, J.
In:

Plant Cell reports

Bandangabe: 28
Auflage: 7
ISBN: ISSN 0721-7714
Seite: 1075 - 1083
Jahr: 2009

Einordung:
Institut: Professur Agrobiotechnologie

Abstract:
We have developed a self-excision Cre-vector
to remove marker genes from Brassica napus. In this vector cre recombinase gene and bar expression cassette were inserted between two lox sites in direct orientation. These lox-flanked sequences were placed between the seed-specific
napin promoter and the gene of interest (vstI). Tissuespecific cre activation resulted in simultaneous excision of the recombinase and marker genes. The vector was introduced into B. napus by Agrobacterium-mediated transformation.
F1 progeny of seven lines with single and multiple
transgene insertions was subjected to segregation and molecular analysis. Marker-free plants could be detected and confirmed by PCR and Southern blot in all transgenic lines tested. The recombination efficiency expressed as a ratio of plants with complete gene excision to the total number of investigated plants varied from 13 to 81%
dependent on the transgene copy number. Potential application of this system would be the establishment of markerfree transgenic plants in generatively propagated species.

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