Brazilian Archives of Biology and Technology
Print version ISSN 1516-8913
LOPES, Francis Julio Fagundes et al. Restriction enzyme improves the efficiency of genetic transformations in Moniliophthora perniciosa, the causal agent of witches broom disease in Theobroma cacao. Braz. arch. biol. technol. [online]. 2008, vol.51, n.1, pp. 27-34. ISSN 1516-8913. http://dx.doi.org/10.1590/S1516-89132008000100004.
The presence of restriction enzymes in the transformation mixture improved the efficiency of transformation in Moniliophthora perniciosa. The influence of the vector shape (linear or circular), the patterns of plasmid integration in genomic sites and the influence of the promoter used to express the gene marker were also analyzed. The addition of BamHI or NotI increased the number of transformants by 3-10-fold and 3-fold, respectively, over the control without added enzyme. The use of pre-linearized plasmid did not increase the transformation efficiency in comparison with the circular plasmid. However, the frequency of multi-copy transformants increased significantly. The transformation procedure here reported resulted in better production of protoplasts and transformation efficiency. In addition, the time necessary for the detection of the first transformants and the number of insertions were reduced.
Keywords : Moniliophthora perniciosa; transformation; restriction enzyme.