Alternative of antibiotic selection

[bimal]

I am looking for a simple selection marker other than antibiotic or herbicides/tellurite/arsenite/mercuric salts, because present concept of research is that antibiotics or other selection molecules are stress signalling molecules. So I want some non stress and non signal molecule to perform some transposition experiment in bacteria without any signalling bias. Like, I am thinking a energy source molecule (e.g., carbon source molecule) which e. coli cannot use due to lack of initial metabolic enzyme but may be downstream genes are already present, so in presence of foreign initial metabolic gene in the transposon element it will be able to grow in that particular energy source.

 

Expert opinion or suggestion is highly appreciated.

[CharonY]

Well, in transposons quite often glucuronidase or galactosidase activity are often used. Positive selection is usually done with e.g. X-gal or X-gluc plates.

[bimal]

Dear Charon,

Thank you for your suggestion. Actually, I will do random mutagenesis and I don't want to pick-up colony. Insate of selecting single colony I will directly go to NGS isolating DNA from all the cell from the culture. So, I am looking for some selection where in presence of selection pressure only transposed cell will go.

 

Regards,

Bimal