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bug2012 bug2012
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11 years ago
Predict what would happen after transforming competent E. coli cells with a plasmid without an ori. Would it be possible to use antibiotic selection to identify bacterial colonies containing this plasmid? Briefly explain why or why not.

This question is stumping me. If there's no origin of replication, can the E. coli replicate? If they can't replicate, then you can't test colonies with an antibiotic because the E. coli containing the resistance plasmid would not replicate. Is that right?
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wrote...
11 years ago
The E. Coli will replicate, but the plasmid containing your antibiotic resistance gene would remain in one of the two daughter cells without being replicated. So basically, your population of transformed cells will remain constant while your wild-type cells continue to grow in number.

Bearing this in mind, it's impossible to "grow" resistant colonies under these conditions because the size never changes. Selection experiments are therefore difficult to impossible, unless you succeed in finding your resistant bacteria under a microscope!
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