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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Chapter 29, Problem 6P
Interpretation Introduction
(a)
Interpretation:
The speed of the template DNA that spins at an E.coli replication fork needs to be determined.
Concept introduction:
DNA exists inside the nucleus as a molecule which is double-stranded. One strand is the coding strand while the second strand of DNA is a non-coding strand. The non-coding strand is actually the template (Template DNA).
Interpretation Introduction
(b)
Interpretation:
The relative velocity of movement of DNA polymerase III holoenzyme and template needs to be determined.
Concept introduction:
The DNA polymerase III holoenzyme is the type of primary enzyme complex which is involved in prokaryotic
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Students have asked these similar questions
(a) How fast does template DNA spin (expressed in revolutions per second) at an E. coli replication fork? (b) What is the velocity of movement (in micrometers per second) of DNA polymerase III holoenzyme relative tothe template?
DNA ligase has the ability to relax supercoiled circular DNA in the
presence of AMP but not in its absence.
(a) What is the mechanism of this reaction, and why is it dependent
on AMP?
(b) How might one determine that supercoiled DNA had in fact
been relaxed?
1a.
What do DNA polymerases need to be able to synthesize a new strand of DNA?
In 1970, Fred Sanger and colleagues published a DNA sequencing procedure based on the principles of DNA replication. This
procedure uses in vitro DNA synthesis in the presence of radioactive nucleotides and specific chain-terminators. These specific
chain terminators lack a 3' hydroxyl group and are called 2', 3' – dideoxyribonucleoside triphosphates. This means that once a
chain terminator is built into the newly synthesized strand, no further synthesis can occur on that particular strand. They are most
usually labelled with radioactive 355 (isotope 35 of sulphur). Four reactions are assembled, each containing one each of 2', 3' -
dideoxythymidine triphosphate (ddTTP), ddCTP, ddATP or ddGTP. In each reaction tube, all the fragments will end with same base
(T, C, A or G). These reactions are each loaded in their own lane and separated by gel electrophoresis, exposed to autoradiograph
film (X-ray film) and…
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- Consider the proteins involved in DNA replication. A. What can a regular DNA polymerase do? Choose one or more. i Catalyze formation of a phosphodiester bond between one dNTP and another dNTP ii Catalyze formation of a phosphodiester bond between one DNA fragment and another iii Catalyze formation of a phosphodiester bond between the 3' end of a DNA fragment, and a dNTP iv Catalyze formation of a phosphodiester bond between the 5' end of a fragment and a dNTP o Open up double-stranded DNA to expose the two separate template strands B. For any ONE action DNA polymerase canNOT do, from the list above, name an enzyme that does that action or makes it unnecessary. Enzyme: What action it does, or makes unnecessary (by numbers given above in part a): i ==>> iv Varrow_forwardMolecules of DNA Polymerase III per Cell vs. Growth Rate It is estimated that there are 40 molecules of DNA polymerase III per E. coli cell, is it likely that the growth rate of E. coli is limited by DNA polymerase III availability?arrow_forwardHuman Genome Replication Rate Assume DNA replication proceeds at a rate of 100 base pairs per second in human cells and origins of replication occur every 300 kbp. Assume also that human DNA polymerases are highly processive and only two molecules of DNA polymerase arc needed per replication fork. How long would it take to replicate the entire diploid human genome? How many molecules of DNA polymerase does each cell need to carry out this task?arrow_forward
- Multiple Replication Forks in E. coli II On the basis of Figure 28.2, draw a simple diagram illustrating replication of the circular E. coli chromosome (a) at an early stage, (b) when one-third completed, (c) when two-thirds completed, and (d) when almost finished, assuming the initiation of replication at oriC has occurred only once. Then, draw a diagram showing the E. coli chromosome in problem 3 where the E. coli cell is dividing every 20 minutes.arrow_forwardMultiple Replication Forks in E. coli I Assuming DNA replication proceeds at a rate of 750 base pairs per second, calculate how long it will take to replicate the entire E. coli genome. Under optimal conditions, E. coli cells divide every 20 minutes. What is the minimal number of replication forks per E. coli chromosome in order to sustain such a rate of cell division?arrow_forwardIn the DNA extraction. What is the role of alcohol in the DNA extraction process?arrow_forward
- Show all work. 1. A) Produce a double-stranded piece of DNA that is 11 base pairs long using the letters A, C, G, T and label the ends of both strands using 3' and 5' appropriately. B) Calculate the ratio of (A+T)/(G+C) and (A+G)/(C+T). Explain why one ratio will always be equal to 1.0?arrow_forwardA. DNA Replication Construct a DNA with 15 base pairs. (Note that the first three nucleofides of the parent DNA (3' to 5') strand correspond to a start codon and its last three nucleotides correspond to a stop codon in its MRNA counterpart later on.) Write it down as follows: a. the sequence of parent DNA (template) 3' A C A TT 5' 3' Upon undergoing DNA replication, show what one daughter DNA molecule will look like. Write it down as follows: b. the sequence of DNA Daughter 1: 3' 5' 5' 3' C. the sequence of DNA Daughter 2: 3' 3' 5' in inarrow_forwardI. What is the correct order of enzyme action during DNA replication? Number the steps from 1 to 7. HINT: Refer to the slide show and video lecture on this topic to help you solve this one: Synthesis of RNA primers (priming) Ligation II. A double-stranded DNA molecule with the sequence shown below can produce a polypeptide that is four amino acids long. Identify which DNA strands are the coding and the transcribed template strands by circling C or T to the left of the table below, respectively. Use an arrow to indicate the direction of transcription. In the table, show the mRNA sequences and amino acids in this peptide. In spaces to the left and right of the table, label all 5' and 3' ends of all relevant nucleic acid strands. READ CAREFULLY: The table gives you the possibility of filling in answers that show transcription from either strand or in either direction. You are only required to fill in the information relevant to ONE PEPTIDE (no others). Refer to the genetic code on the…arrow_forward
- Which statements are true? Explain why or why not.1 The different cells in your body rarely havegenomes with the identical nucleotide sequence.2 In E. coli, where the replication fork travels at 500nucleotide pairs per second, the DNA ahead of the fork—in the absence of topoisomerase—would have to rotate atnearly 3000 revolutions per minute.3 In a replication bubble, the same parental DNAstrand serves as the template strand for leading-strandsynthesis in one replication fork and as the template forlagging-strand synthesis in the other fork.4 When bidirectional replication forks from adja-cent origins meet, a leading strand always runs into a lag-ging strand.5 DNA repair mechanisms all depend on the exis-tence of two copies of the genetic information, one in eachof the two homologous chromosomesarrow_forwardPstl. 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.arrow_forwardMutation: Thiamine Dimers A. what is a mutagen or cellular process that leads to this mutation? B. If the nucleotide excision repair pathway is not functional, is there another pathway or mechanism for preparing this mutation? if so, what is the pathway/mechanism? C. what is the consequence of this mutation if it is not repaired before DNA is replicated to make the next generation of cells?arrow_forward
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