Biochemistry
9th Edition
ISBN: 9781319114671
Author: Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher: W. H. Freeman
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Question
Chapter 29, Problem 8P
Interpretation Introduction
(a)
Interpretation:
From the given twist number and writhe number, the linking number is to be calculated .
Concept introduction:
Circular DNA generally undergoes super coiling in order to fit itself inside a smaller space like a cell. Plasmids are the number of extra chromosomal material found inside bacterial cells. They are generally found in super coiled state.
Interpretation Introduction
(b)
Interpretation:
The writhe number is to be calculated based on the twist number and the linking number.
Concept introduction:
Circular DNA generally undergoes super coiling in order to fit itself inside a smaller space like a cell. Plasmids are the number of extra chromosomal material found inside bacterial cells. They are generally found in super coiled state.
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Pick a plasmid . What was its approximate transformation? Express it in # colonies per microgram of DNA transformed.
Assume the original DNA was about .001 ug/ul . Count how many colonies you got on one plate (or estimate that number) and figure out how much of the total solution you plated on that plate. Multiply by all the plates, if you plated all of it. OR, if you only plated some of it, figure out how many colonies you would have gotten had you plated all of it. Divide by the number of ug used.
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As this is a non-directional cloning, recombinant plasmids can contain an insert ligated into the vector in two different orientations. Provide two diagrams to illustrate the two potential recombinant plasmids, with the inserts ligated in opposite orientations. Include all RE sites and distances between sites on the diagram.
Pstl.
EcoRI
Origin of
replication
(ori)
Ampicillin Tetracycline
resistance resistance
(Amp) (Tet")
pBR322
(4,361 bp)
BamHI
Pvull
Sall
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Plasmid DNA
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No plasmid DNA
pBR322 (no insert)
Recombinant plasmid
00,000.
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Transformation
of E. coli cells
+AMP plate
pBR322
Figure 7-5
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Based on the recombinant plasmid growth pattern (bottom row of blue table), which of
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- Plasmid Mapping (what is it? why is it important?).arrow_forwardYou have another circular plasmid. Complete and effective digestion of this plasmid with a restriction enzyme yields three bands: 4kb, 2kb, and 1 kb. In comparing the band intensity on an ethidium bromide-stained gel, you notice that the 4 kb and the 2 kb bands have the exact same brightness. The 1 kb band is exactly one fourth as bright as each of these. (Assume there is uniform staining with ethidium bromide throughout the gel.) How many times did the enzyme cut the plasmid? What is the size of the plasmid? Justify your answers to a and b above using a clearly labeled diagram showing the relative location of the cut-sites on the plasmid.arrow_forwardPlease answer this asap. Thanks, You have discovered a new plasmid RK21 in a unique bacterial community. As a first step towardunderstanding this plasmid, you digest the plasmid with three restriction enzymes: SspI, XhoI andSmaI. You run the digested plasmid DNA on an agarose gel, along with an uncut sample of theRK21 plasmid DNA as a control.Unfortunately you forget to load a DNA ladder, and obtain the following results. Assumecomplete digestion of all samples or all the digests worked completelyarrow_forward
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