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A researcher ligates a gene of interest into the EcoR I restriction site of the cloning vector pBR322 and introduces the resulting mixture into competent E. coli cells. Which of the following statements regarding this experiment is incorrect?

Options

  1. AThe recombinant plasmid confers resistance to both ampicillin and tetracycline.
  2. BRecombinant transformants can be distinguished from non-recombinants by replica plating on antibiotic-containi
  3. CThe EcoR I recognition sequence is located outside the selectable marker genes in pBR322.
  4. DBoth recombinant and non-recombinant transformants will form colonies on a medium containing ampicillin.

Correct answer

B. Recombinant transformants can be distinguished from non-recombinants by replica plating on antibiotic-containi

Step-by-step solution

In the cloning vector pBR322, the EcoR I restriction site is located outside both the ampicillin resistance and tetracycline resistance genes. Therefore, insertion of an alien DNA at the EcoR I site does not cause insertional inactivation of either selectable marker. Both recombinant and non-recombinant plasmids will continue to confer resistance to ampicillin and tetracycline. Because their antibiotic resistance profiles are identical, replica plating on antibiotic-containing media cannot be used to distinguish re

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