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.

中文翻译：

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DNA 水印：概念证明。

背景 基于 DNA 的水印是识别未经授权使用受专利保护的转基因生物 (GMO) 的有用工具。计算机分析表明，在编码区，同义密码子可用于通过使用 DNA-Crypt 算法将加密信息插入生物体的基因组中。结果 我们在 Vam7 序列中集成了一个认证水印。在我们的研究中，我们使用了一种名为 CG783 的突变酿酒酵母菌株，它在 Vam7 序列中具有琥珀色突变。CG783 细胞不能形成孢子，此外还显示出异常的液泡形态。用 pRS314 Vam7 转化 CG783 产生与野生型酵母菌株 CG781 非常相似的表型。整合的水印不影响 Vam7 的功能，并且与用 pRS314 Vam7 转化的 CG783 细胞相比，用 pRS314 Vam7-TB 转化的 CG783 细胞的所得表型没有显着差异。结论 从我们的实验中我们得出结论，DNA-Crypt 产生的 DNA 水印不影响从 mRNA 到蛋白质的翻译。通过分析液泡形态、生长速率和孢子形成能力，我们证实所得 Vam7 蛋白具有功能活性。