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 5, Problem 3P
Interpretation Introduction
Interpretation:
The average distance between the cleave sites for the given enzymes AluI and NotI on digestion of double stranded DNA is to be stated.
Concept introduction:
The restriction enzymes are generated mostly by the bacteria. In the formation of recombinant DNA, the restriction enzymes are involved to cut the particular region in the DNA molecule. This region is known as restriction site.
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Pstl.
EcoRI
Origin of
replication
(ori)
Ampicillin Tetracycline
resistance resistance
(Amp) (Tet")
pBR322
(4,361 bp)
BamHI
Pvull
Sall
Recombinant
Plasmid DNA
Bacterial cell contains...
No plasmid DNA
pBR322 (no insert)
Recombinant plasmid
00,000.
Host DNA
Transformation
of E. coli cells
+AMP plate
pBR322
Figure 7-5
+TET plate
Based on the recombinant plasmid growth pattern (bottom row of blue table), which of
the depicted plasmid's restriction sites was used to prepare this sample? Explain how
you can tell.
B PLEASE second one
Restriction sites of Lambda (A) DNA - In base pairs (bp)
The sites at which each of the 3 different enzymes will cut the same strand of lambda DNA
are shown in the maps (see figure 3 B-D), each vertical line on the map is where the respective
enzymes will cut.
A DNA
A
(bp)
48502
10 000
20 000
30 000
40 000
9162
17 198
B
Sal I
7059
14 885
28 338
35 603
42 900
(bp)
Hae III
11 826
21 935
29 341
38 016
(bp)
11648
29,624
Eco R1
(bp)
10 592 16 246
28 915
41 864
Figure 3: Restrictrion site map showing the following A) inear DNA that is not cut as reference B) DNA CLt with Sal L C) DNA cut with Hae , D)
DNA cut with Eco RI
1. Calculate the size of the resulting fragments as they will occur after digestion and write
the sizes on the maps below. Note that linear DNA has a total size of 48 502 bp (see
figure 3A).
Page 3 of 7
9162
17 198
Sal i
(bp)
7059
14 885
28 338
35 603
42 900
Hae I
(bp)
11 826
21 935
29 341
38 016
11648
29,624
Eco R1
(bp)
10 592
16 246
28 915
41 864
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