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DNA watermarks: a proof of concept.

Heider D, Barnekow A - BMC Mol. Biol. (2008)

Bottom Line: The integrated watermark did not influence the function of Vam7 and the resulting phenotype of the CG783 cells transformed with pRS314 Vam7-TB shows no significant differences compared to the CG783 cells transformed with pRS314 Vam7.From our experiments we conclude that the DNA watermarks produced by DNA-Crypt do not influence the translation from mRNA into protein.By analyzing the vacuolar morphology, growth rate and ability to sporulate we confirmed that the resulting Vam7 protein was functionally active.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Experimental Tumorbiology, University of Muenster, Badestrasse 9, 48149 Muenster, Germany. dominik.heider@uni-muenster.de

ABSTRACT

Background: DNA-based watermarks are helpful tools to identify the unauthorized use of genetically modified organisms (GMOs) protected by patents. In silico analyses showed that in coding regions synonymous codons can be used to insert encrypted information into the genome of living organisms by using the DNA-Crypt algorithm.

Results: We integrated an authenticating watermark in the Vam7 sequence. For our investigations we used a mutant Saccharomyces cerevisiae strain, called CG783, which has an amber mutation within the Vam7 sequence. The CG783 cells are unable to sporulate and in addition display an abnormal vacuolar morphology. Transformation of CG783 with pRS314 Vam7 leads to a phenotype very similar to the wildtype yeast strain CG781. The integrated watermark did not influence the function of Vam7 and the resulting phenotype of the CG783 cells transformed with pRS314 Vam7-TB shows no significant differences compared to the CG783 cells transformed with pRS314 Vam7.

Conclusion: From our experiments we conclude that the DNA watermarks produced by DNA-Crypt do not influence the translation from mRNA into protein. By analyzing the vacuolar morphology, growth rate and ability to sporulate we confirmed that the resulting Vam7 protein was functionally active.

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Sporulation morphology. A: Sporulation of the CG781 yeast strain; B: the sporulation deficient CG783 yeast strain; C: sporulation of the pRS314 Vam7 transformed CG783 yeast strain; D: sporulation of the pRS314 Vam7-TB transformed CG783 yeast strain (magnification 100×).
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Figure 6: Sporulation morphology. A: Sporulation of the CG781 yeast strain; B: the sporulation deficient CG783 yeast strain; C: sporulation of the pRS314 Vam7 transformed CG783 yeast strain; D: sporulation of the pRS314 Vam7-TB transformed CG783 yeast strain (magnification 100×).

Mentions: In contrast to strain CG783, which is not able to sporulate most likely due to the lack of a functionally active Vam7 in CG783 cells, 72.4% ± 2.19 of the pRS314 Vam7 and 72.2% ± 2.59 of the pRS314 Vam7-TB transformed yeast cells formed spores. The spores of the transformed CG783 cells displayed the normal phenotype of CG781 spores (Figures 6, 7). Also in these experiments the insertion of the watermark did not result in an altered phenotype, comparing CG783 cells transformed with pRS314 Vam7 and CG783 cells transformed with pRS314 Vam7-TB.


DNA watermarks: a proof of concept.

Heider D, Barnekow A - BMC Mol. Biol. (2008)

Sporulation morphology. A: Sporulation of the CG781 yeast strain; B: the sporulation deficient CG783 yeast strain; C: sporulation of the pRS314 Vam7 transformed CG783 yeast strain; D: sporulation of the pRS314 Vam7-TB transformed CG783 yeast strain (magnification 100×).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2375902&req=5

Figure 6: Sporulation morphology. A: Sporulation of the CG781 yeast strain; B: the sporulation deficient CG783 yeast strain; C: sporulation of the pRS314 Vam7 transformed CG783 yeast strain; D: sporulation of the pRS314 Vam7-TB transformed CG783 yeast strain (magnification 100×).
Mentions: In contrast to strain CG783, which is not able to sporulate most likely due to the lack of a functionally active Vam7 in CG783 cells, 72.4% ± 2.19 of the pRS314 Vam7 and 72.2% ± 2.59 of the pRS314 Vam7-TB transformed yeast cells formed spores. The spores of the transformed CG783 cells displayed the normal phenotype of CG781 spores (Figures 6, 7). Also in these experiments the insertion of the watermark did not result in an altered phenotype, comparing CG783 cells transformed with pRS314 Vam7 and CG783 cells transformed with pRS314 Vam7-TB.

Bottom Line: The integrated watermark did not influence the function of Vam7 and the resulting phenotype of the CG783 cells transformed with pRS314 Vam7-TB shows no significant differences compared to the CG783 cells transformed with pRS314 Vam7.From our experiments we conclude that the DNA watermarks produced by DNA-Crypt do not influence the translation from mRNA into protein.By analyzing the vacuolar morphology, growth rate and ability to sporulate we confirmed that the resulting Vam7 protein was functionally active.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Experimental Tumorbiology, University of Muenster, Badestrasse 9, 48149 Muenster, Germany. dominik.heider@uni-muenster.de

ABSTRACT

Background: DNA-based watermarks are helpful tools to identify the unauthorized use of genetically modified organisms (GMOs) protected by patents. In silico analyses showed that in coding regions synonymous codons can be used to insert encrypted information into the genome of living organisms by using the DNA-Crypt algorithm.

Results: We integrated an authenticating watermark in the Vam7 sequence. For our investigations we used a mutant Saccharomyces cerevisiae strain, called CG783, which has an amber mutation within the Vam7 sequence. The CG783 cells are unable to sporulate and in addition display an abnormal vacuolar morphology. Transformation of CG783 with pRS314 Vam7 leads to a phenotype very similar to the wildtype yeast strain CG781. The integrated watermark did not influence the function of Vam7 and the resulting phenotype of the CG783 cells transformed with pRS314 Vam7-TB shows no significant differences compared to the CG783 cells transformed with pRS314 Vam7.

Conclusion: From our experiments we conclude that the DNA watermarks produced by DNA-Crypt do not influence the translation from mRNA into protein. By analyzing the vacuolar morphology, growth rate and ability to sporulate we confirmed that the resulting Vam7 protein was functionally active.

Show MeSH
Related in: MedlinePlus