A circular bacterial plasmid containing a gene for tetracycline resistance was c

Question

A circular bacterial plasmid containing a gene for tetracycline resistance was cut with restriction enzyme BglII. Electrophoresis showed one band of 14 kb.The plasmid was cut with EcoRV and electrophoresis produced two bands, one of 2.5 kb and the other 11.5 kb.

Plasmid DNA cut with BglII was mixed and ligated with donor DNA fragments, also cut with BglII, to make recombinant DNA molecules. All recombinant clones proved to be tetracycline sensitive.

d. Give the most likely reason why the recombinant molecules lost their tetracycline resistance capabilities.

Explanation / Answer

A circular bacterial plasmid containing a gene for tetracycline resistance was cut with restriction enzyme BglII. Electrophoresis showed one band of 14 kb.The plasmid was cut with EcoRV and electrophoresis produced two bands, one of 2.5 kb and the other 11.5 kb.

Plasmid DNA cut with BglII was mixed and ligated with donor DNA fragments, also cut with BglII, to make recombinant DNA molecules. All recombinant clones proved to be tetracycline sensitive.

d. recombinant molecules lost their tetracycline resistance capabilities because the DNA fragment was inserted in the middle of the gene cluster conferring resistance to tetracyclinetetR.

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