I cultured transformed BL21DE3 in LB broth from colonies. The colonies were pure white in color. The cells were transformed with my gene of interest. But the culture turned into pink. I have added proper antibiotic also. Is there any contamination? What could be the possible reasons?
Dear Prabhash Mahanta
The colour is quite stange.
Are you sure that the colour is not related to the expression of your gene of interest? It is a metallo protein? o sometingh that can justify the formation of that colour?
best regards
Manuele
Thanks for responding Manuele Martinelli
Yes, it is a metalloprotein. It consists one Zinc atom. I don’t have any idea from where the color arised. I did subculture from the colonies(from the same plate) earlier also. But it didn’t look like this.
It may be your plasmid, but I would suggest you streak out from the culture on a plain LB plate (not an amp plate) and be sure you don't have some other contaminating bacteria. If all the colonies that grow appear homogenous then you will be comfortable assuming it is the protein you are making.
It could also be a fungus, some yeast grows pink. I agree with the idea of a re-streak and colony morphology check. If you do have a mix, you should be able to re-isolate your desired bacteria (be sure to use the amp in plates for that process or you're likely to lose the plasmid).
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