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Upon selection in puromycin, it is probable that the non-integrated allele will be mutated due to indels created by imperfect NHEJ, hence the tagged protein will remain the only functional copy within the cell, which also enables the functionality of the tagged protein to be tested.
Meanwhile, we suggest that there is at least one additional molecular target in the R. opacus genome which fully enables the functionality of xylA and xylB genes to generate the xylose-fermenting strain capable of efficiently producing TAGs at high xylose concentrations.
We further demonstrated that in addition to the introduction and the concomitant expression of heterologous xylA and xylB genes, there is another molecular target in the R. opacus genome which fully enables the functionality of xylA and xylB genes to generate the robust xylose-fermenting strain capable of efficiently producing TAGs at high xylose concentrations.
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Characterizing regions that are affected by selection may enable inferences on the functionality of a genomic region and possibly the effects of specific genes or gene combinations on specific traits [ 3- 6].
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