I am on gene work to express gene of interest in some Bacillus species. Thus far, i need to try some RBS in Vector which is optimized for bacillus subtilis.
Before i design vector to express in other species , I have transformed subtilis vector in other species and then i could select transformants by antibiotic selection
I have known GoI cannot be expressed without RBS and I have to insert RBS depends on bacteria strain individually.
so i wondered if there's no working RBS then Resistant gene in vector also cannot work???
Thank you in advance!!
Translational efficiency is not a binary yes or no. It is a gradient. The quality of ribosome recognition of the start codon varies with the quality of the RBS but is usually never zero. So even without a good RBS you probably will have some translation occurring. Is this sufficient for selection? That can only be determined empirically by experiment.
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