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

作者：Kopertekh Lilya; Broer Inge; Schiemann Joachim^*

来源：Plant Cell Reports, 2009, 28(7): 1075-1083.

DOI：10.1007/s00299-009-0711-5

• Brassica napus
• Biosafety
• Marker gene
• Self-excision Cre-vector
• Seed-specific napin promoter

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). Tissue-specific 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 marker-free transgenic plants in generatively propagated species.

• 出版日期2009-7