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Standard VII
Biology
Question

From the below list, which of the following is the most logical sequence of steps for splicing foreign DNA into a plasmid and inserting the plasmid into a bacterium?
(i) Transformbacteria with recombinant DNA molecule.
(ii) Cut the plasmid DNA using restriction enzymes.
(iii) Extract plasmid DNA from bacterial cells.
(iv) Hydrogen-bond the plasmid DNA to nonplasmid DNA fragments.
(v) Use ligase to seal plasmid DNA to nonplasmid DNA.
  1. (iv), (v), (i), (ii), (iii)
  2. (iii), (ii), (iv), (v), (i)
  3. (iii), (iv), (v), (i), (ii)
  4. (ii), (iii), (v), (iv), (i)

A
(iii), (ii), (iv), (v), (i)
B
(iii), (iv), (v), (i), (ii)
C
(ii), (iii), (v), (iv), (i)
D
(iv), (v), (i), (ii), (iii)
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Solution
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1. A gene of interest ( DNA fragment) is isolated from cells that have been grown in laboratory culture.
2. Both the human DNA and the plasmid are treated with the same restriction enzyme to produce identical sticky ends.
3. The restriction enzyme cuts the plasmid DNA at its single recognition sequence, disrupting the antibiotic resistance gene.
4. The DNA fragment are mixed together and the complementary sticky ends are attached to each other by base pairing. The enzyme DNA- ligase is added to bond the sticky ends.
5. The recombinant plasmid or molecular clone, is introduced into a bacterial cell by adding the DNA to a bacterial culture. Under the right condition some bacteria will take up the plasmid from solution by the process transformation.
Therefore, the correct answer is option B.

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Q1
From the below list, which of the following is the most logical sequence of steps for splicing foreign DNA into a plasmid and inserting the plasmid into a bacterium?
(i) Transformbacteria with recombinant DNA molecule.
(ii) Cut the plasmid DNA using restriction enzymes.
(iii) Extract plasmid DNA from bacterial cells.
(iv) Hydrogen-bond the plasmid DNA to nonplasmid DNA fragments.
(v) Use ligase to seal plasmid DNA to nonplasmid DNA.
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Q2
Identify the correct sequence of steps in the construction of recombinant DNA.
(1) use restriction enzymes
(2) use DNA ligase
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(4) introduce plasmid into new host bacterium.
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Q3
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I. Complementary sticky ends are produced on the foreign DNA and the plasmid.
II. Foreign DNA and plasmids are mixed together
III. DNA ligase is used to seal the nicks in the sugar–phosphate backbone.
IV. Cut the foreign DNA (containing the DNA fragment of interest) and the plasmid with the same restriction enzyme.
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Q4
A foreign DNA and plasmid cut by the same restriction endonuclease can be joined to form recombinant plasmid using

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Q5
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