A Graduate Student Wants to Clone a Particular Gene into a Plasmid.The

question 4

Multiple Choice

A graduate student wants to clone a particular gene into a plasmid.The sequence includes AluI and BamHI sites on both sides of the desired fragment.AluI cuts symmetrically directly between the G and C nucleotides in a palindromic 5' AGCT 3' sequence.BamHI cuts asymmetrically directly between the G and G nucleotides in a palindromic5' GGATCC 3' sequence.Which of the two restriction endonucleases should the graduate student choose,and why?

• A)BamHI to cut both sides-since it cuts asymmetrically,it'll leave the sticky,cohesive single-strand DNA ends that will make it easier to ligate into a BamHI-cut plasmid DNA sequence.
• B)AluI to cut both sides-it's always easier to ligate together blunt ends of DNA.She should also use AluI on the plasmid she wants to put the fragment into.
• C)BamHI on the fragment,and AluI on the plasmid-this will give her the matching sequences to anneal/ligate together on the fragment/plasmid combination.
• D)BamHI on one side of the fragment,and AluI on the other side-this would keep the fragment from sticking right back to where it was cut out from in the original DNA.

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