Is it possible that two different restriction enzymes couldcut the human genome into exactly the same number offragments and with exactly the same distribution of fragment sizes, yet the ends produced by the two enzymescould not be joined together by DNA ligase? Explain

Is it possible that two different restriction enzymes couldcut the human genome into exactly the same number offragments and with exactly the same distribution of fragment sizes, yet the ends produced by the two enzymescould not be joined together by DNA ligase? Explain

Biology Today and Tomorrow without Physiology (MindTap Course List)

5th Edition

ISBN:9781305117396

Author:Cecie Starr, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Christine Evers, Lisa Starr

Chapter10: Biotechnology

Section: Chapter Questions

Problem 1CT

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Concept explainers

Genetic Evolution

Sexual reproduction produces distinct population variations. Heredity determines the traits and qualities to be passed down to the progeny. Over time, this variation within-species is also affected by the process of human evolution. The study of how genetic variation leads to evolutionary changes is known as genetic evolution. If a trait helps an organism to survive and reproduce, it is more likely to be passed down the generations. It is rare that such traits result in a completely distinct set of traits in descendants than the ancestral population; this can result in the emergence of a new species.

Question

Is it possible that two different restriction enzymes could
cut the human genome into exactly the same number of
fragments and with exactly the same distribution of fragment sizes, yet the ends produced by the two enzymes
could not be joined together by DNA ligase? Explain

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During PCR amplification in preparation for DNA sequencing, why were there different colors at the 3’ ends of the fragments produced? (What did these four colors represent?)
If the GAATTC palindrome is repeated four times on the same piece of linear DNA, and the restriction enzyme that recognizes that base sequence is present and digests the DNA, how many DNA fragments will be produced?
In making recombinant DNA molecules that combine restriction fragments from different organisms, researchers usually prefer restriction enzymes like BamHI or HindIII that generate fragments with “sticky ends” (ends with overhangs) rather than enzymes like HpaI or SmaI (Table 12.1) that generate fragments with “blunt ends” (ends without overhangs). Can you think of a reason for this preference?
In making recombinant DNA, what is the benefit of using a restriction enzyme that cuts DNA in a staggered fashion?
Consider a partial restriction digestion, in which genomic DNA is exposed to a small, limiting amount ofa restriction enzyme for a very short period of time.a. Would the resultant fragments be longer or shorteror the same size as those produced by a completedigestion?b. If you prepared genomic DNA from a tissue sample containing millions of cells, would the fragments produced by partial digestion of DNA fromall of these cells be the same or different?
When linear DNA is sequenced, the nucleotide base pairs are numbered from the start to finish. a DNA molecule that is 3133 base pairs long is digested with RsaI restriction enzyme recognition sites at base numbers 366, 1534, and 2207. What are the sizes of the DNA fragments that will be produced after the DNA is digested with RsaI?
Above are the results of gel electrophoresis following digestion with restriction enzymes. What is the total length of the DNA fragment? Which enzyme, HindIII or EcoRI, produced a larger fragment? What part of DNA causes it to be negatively charged in order for electrophoresis to work?
Your cloning vector has restriction recognition sites for two restriction endonucleases, EcorI and BamHI. However, the DNA to be manipulated does not have recognition sites for these two restriction endonucleases. How would you construct a recombinant DNA for the given DNA?
What sequence do the restriction enzymes used in the lab recognize? How do they cut? And how would these different cuts effect cloning? (i.e. compare and contrast how overhang and blunt cuts will differ with cloning.) What organisms were the restriction enzymes used in this lab derived from? Why are restriction enzymes important for the organism that makes them? What is the purpose of multiple cloning sites on pUC19? What size were each of your plasmid fragments?
Consider the ends of the DNA fragments shown below. They have been produced by digestion of a single sequence of DNA using a number of restriction endonucleases. 1. 5'A 3' 3'TTCGA5' 2. 5'G 3' 3'CAGCT5' 3. 5'AATTC3' 3' G5 4. 5'TCGAC3' 3' G5' 5. 5'GGG 3' 3'CCC 5' Which of these ends are capable of annealing and being joined by DNA ligase?
How would the results of this activity have been different if the DNA sequence you digested were circular instead of linear? When restriction digests are performed and analyzed in the laboratory, one lane of the agarose gel is loaded with DNA ladders, which are DNA fragments of known sizes. What is the point of including these size standards?
Restriction enzymes and DNA ligase play essential roles in DNA cloning. How is it that a bacterium that produces a restriction enzyme does not cut its own DNA?