Laboratory Experiments in Microbiology (12th Edition) (What's New in Microbiology)
12th Edition
ISBN: 9780134605203
Author: Ted R. Johnson, Christine L. Case
Publisher: PEARSON
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Textbook Question
Chapter 30, Problem 2Q
Using this map of pLAB30 (18 kb), give the number and lengths of the restriction fragments that would result from digesting pLAB30 with EcoR1, with BamHI, and with both enzymes together.
Which enzyme will give the smallest piece containing the tetracycline-resistance gene?
Expert Solution & Answer
Summary Introduction
To determine:
The size and number of DNA Fragments given by restriction enzymes Eco RI, Bam HI, and both these enzymes together.
Introduction:
Restriction enzyme is an enzyme that cuts the DNA fragment at the specific site by recognizing the DNA sequence at that site. The site at which the DNA is cut is termed as the restriction site.
Explanation of Solution
pLAB30 is an 18 kb map, which is cut by two restriction enzymes, Eco RI, and Bam HI. When Eco RI acts, then it cuts the DNA fragment at two sites, giving an 11 kb fragment and 7 kb fragment of the DNA.
When Bam HIacts, then it cuts DNA fragment at three sites, giving three fragments. The size of these fragments is 9 kb, 3 kb, and 6 kb.
When both these restriction enzymes act together, then 5 DNA fragments are obtained. The size of these fragments is 1 kb, 3 kb, 3 kb, 3kb, and 8 kb.
Conclusion
The table showing the size and number of DNA fragments after the DNA is acted upon by restriction enzymes is shown below:
| Restriction enzyme | Number of DNA fragments obtained | Size of DNA fragments |
| Eco RI | 2 | 11 kb and 7 kb |
| Bam HI | 3 | 9 kb, 3 kb, and 6 kb |
| Eco RI + Bam HI | 5 | 1 kb, 3 kb, 3 kb, 3kb, and 8 kb |
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For Pet41 (choose Pet41 a, b, or c as provided in the image) how would you design the primers (forward and reverse) for the following gene of interest and what restriction enzymes would be used (as shown in the image)? Be sure to explain and elaborate on why selected and how.
Gene of Interest:
atgggc gacaaaggga 241 cccgagtgtt caagaaggcc agtccaaatg gaaagctcac cgtctacctg ggaaagcggg 301 actttgtgga ccacatcgac ctcgtggacc ctgtggatgg tgtggtcctg gtggatcctg 361 agtatctcaa agagcggaga gtctatgtga cgctgacctg cgccttccgc tatggccggg 421 aggacctgga tgtcctgggc ctgacctttc gcaaggacct gtttgtggcc aacgtacagt 481 cgttcccacc ggcccccgag gacaagaagc ccctgacgcg gctgcaggaa cgcctcatca 541 agaagctggg cgagcacgct taccctttca cctttgagat ccctccaaac cttccatgtt 601 ctgtgacact gcagccgggg cccgaagaca cggggaaggc ttgcggtgtg gactatgaag 661 tcaaagcctt ctgcgcggag aatttggagg agaagatcca caagcggaat tctgtgcgtc 721 tggtcatccg gaaggttcag tatgccccag agaggcctgg cccccagccc acagccgaga 781 ccaccaggca gttcctcatg tcggacaagc ccttgcacct…
If the sequence of base pairs along a DNA molecule occurs strictly at random, what is the expected frequency of a specific restriction enzyme recognition sequence of length (a) four and (b) six base pairs?
Refer to the diagram of pUC18 (Fig.) to determine which restriction enzymes you could use to insert a gene that would interfere with production of β-galactosidase by the host cell.
Chapter 30 Solutions
Laboratory Experiments in Microbiology (12th Edition) (What's New in Microbiology)
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