You are using restriction maps in an newly discovered linearVirus genome. Follow
ID: 5806 • Letter: Y
Question
You are using restriction maps in an newly discovered linearVirus genome. Following enzyme digests, the products areseparated by DNA electrophoresis to yield the followingresults. EcorI digests VG to yield 8.6kb, 7.9 kb and 2.3 kbfragments Not1 digest the 8.6 kb fragments to yeild 7.1 and l.5 kb Not1 digest the 7.9 to yield 5 kb and 2.9 kb fragments Not1 digest the 2.3 kb to yield a 2.3 kb fragmentNot1 digest of VG yielded 10 kb, 5 kb and 3.8 kb fragements. Draw a restriction map of the Viral Genome in linear form. You are using restriction maps in an newly discovered linearVirus genome. Following enzyme digests, the products areseparated by DNA electrophoresis to yield the followingresults. EcorI digests VG to yield 8.6kb, 7.9 kb and 2.3 kbfragments Not1 digest the 8.6 kb fragments to yeild 7.1 and l.5 kb Not1 digest the 7.9 to yield 5 kb and 2.9 kb fragments Not1 digest the 2.3 kb to yield a 2.3 kb fragment
Not1 digest of VG yielded 10 kb, 5 kb and 3.8 kb fragements. Draw a restriction map of the Viral Genome in linear form.
Explanation / Answer
Remember that when mapping restriction digests one cut makes twopieces, two cuts make three pieces and so forth. The best way todetermine the restriction sites on a genome is to draw a line andlabel possible positions of the site. The pieces go together like apuzzle and math can verify your answer. The enzyme EcoR1 cut the viral genome into three pieces, so we candeduce this certain portion of the genome had two restriction sitesfor that enzyme. By adding up the kb given from that digest we cansee this molecule of viral DNA is 18.8 kb's long as shown in theequation below. 8.6 kb + 7.9 kb + 2.3 kb = 18.8 kb The enzyme Not1 cut the DNA molecule into three pieces as well, sowe can deduce there are also two restriction sites for that enzyme.We also arrive at the same size DNA molecule when adding up thepieces from this specific digest. To put the puzzle pieces together, first take them apart. The Viral Genome is represented by a line _______________________________________________________ which was cut into three pieces by EcoR1 _______________________________ 8kb(Not1 cut this piece into __________ 7.1kb and ___ 1.5kb) _________________________ 7.9kb (Not1 cut this piece into _______ 5kb and _____ 2.9kb) _______ 2.3kb (Not1 did not cut this piece, so it has no restrictionsite for Not 1) Again the Viral Genome is represented by a line_____________________________________________________ which was cut into three pieces by Not1 ___________________________________ 10kb ________________ 5kb ________ 3.8kb From these facts we can determine the restriction sites of bothenzymes and checking our answer by adding the fragments together tosee if they reflect our digest. The linear restriction map for Viral Genome is hererepresented by a line with periods to denote restriction sites. _______________.__________._______________________________.___.______ (not to scale) Not1 13.8kb EcoR110.9kb Not1 3.8kb EcoR1 2.3kb
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