ZOL 341 Lecture Notes - Lecture 5: Consensus Sequence, Dnab Helicase, Origin Of Replication

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31 Aug 2016

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Proteins (enzymes) the read the dna and assemble the corresponding strand. There are four stages of replication: initiation, unwinding, elongation, termination. Replication enzymes are attracted to the sequence that make up the replication origin: when talking about bacteria, e. coli is commonly referred to, the origin in e. coli is called oric. Similar sequences at the origin are found all bacteria: conserved sequences. Enzymes must locate and attach to the oric consensus sequence in order for replication to occur. Important enzymes: dnaa- breaks the h-bonds between a and t in the sequence, dnab- separate strands and unwind the helix. Is considered a helicase: dnac- carries dnab to the helix, single stranded binding protein- prevents the dna strands from reforming the. Stress is created during the unwinding process: can lead to supercoiling, dna topoisomerase prevents the dna from knotting. This enzyme works by cleaving and repairing the stands.

Related Questions

You decide to identify the CFTR mutation by analyzing the genomic DNA of your patients compared to healthy individuals. You specifically are looking to see whether a specific 3' gene truncation has occurred in the patients. You will determine this using hybridization techniques with samples from healthy and CF patients. Which of the following will allow you to accomplish this?

	Using an RNA probe complementary to the region not removed by the truncation.
	Using an RNA probe complementary to the region removed by the truncation.
	Using an DNA probe complementary to the region not removed by the truncation.
	Using an DNA probe complementary to the region removed by the truncation.

To conduct the hybridization experiment, you are trying to decide between using a DNA or RNA probe. Which would be ideal to use and why?

	As both are composed of nucleic acids, using either would result in identical results.
	An RNA probe because RNA has uracil bases.
	An RNA probe because it could also be used in a translation experiment.
	A DNA probe because it is more stable than RNA.
	A DNA probe because RNA cannot bind to DNA.

Imagine Hershey/Chase had used an RNA virus (genome composed of RNA) instead of a DNA virus in their experiment. Would radioactivity still have been found in the pellet?

	No, because only DNA can be labeled with radioactivity.
	No, because the RNA genome would not enter the bacteria upon infection.
	No, because while DNA and RNA nucleotides are similar, they are not identical.
	Yes, because DNA and RNA nucleotides are similar.
	Yes, because genome in any form (DNA, RNA, protein) would be labeled similarly.

The human genome consists mostly of non-coding DNA. Which of the following are benefits of this?

	Random DNA mutations generally won't affect RNA and protein function.
	It is faster to duplicate the genome when these are present.
	The existence of introns can lead to multiple variations of proteins encoded by a single gene.
	It is unlikely transposons would exist in the genome if there was too much protein coding DNA.

You accidentally add a mutant dNTP (which has an H instead of an OH connected to the 3â carbon) to a reaction where DNA is being replicated. Which of the following is true of this mutant dNTP?

	It can be incorporated into DNA strand but cannot form a phosphodiester bond with an incoming wild type dNTP
	It can be incorporated into a DNA strand but cannot base pair with a complementary nucleotide
	It can be incorporated into a DNA strand and can form a phosphodiester bond with an incoming dNTP, but only if it is another mutant dNTP
	It cannot be incorporated into a DNA strand.

Andrew Murray's sister, Andrea, is adding to her brother's work on chromosomes. She is using cells that are unable to synthesize adenine (âade) and histidine (âhis). The plasmid she is currently working with consists of an origin of replication and the Ade gene.

Following her transformation of the plasmid into her yeast, what media will the cells be plated on to select for cells that have picked up the plasmid?

	Media containing histidine
	Media containing adenine
	Media lacking adenine
	Media lacking histidine

She starts by attempting to add the centromere DNA into a plasmid containing the origin of replication. Unfortunately, when adding the centromere sequence, the origin of replication is removed, thus leaving a plasmid with only a centromere and selection marker. Following plasmid transformation, what growth result will she see on her plates?

	No colonies
	Little colonies
	Big colonies

She fixes the mistakes from the previous experiment and now has a complete plasmid (selection marker, origin of replication, centromere). She then inserts telomere sequences into the plasmid. How will this impact her transformation?

	It will not impact her transformation
	Her transformation will no longer work because plasmids donât require telomeres
	She will now see much larger colonies
	She will now see fewer but larger colonies
	She will now see smaller colonies

In the Meselson/Stahl experiment, E. coli were first grown in media containing heavy nitrogen, 15N, and then transferred to light nitrogen, 14N, at the beginning of the experiment.

Imagine that their data showed that replication occurs in a conservative manner instead of semi-conservative. What fraction of the DNA helices will consist of mixed DNA after 4 rounds of replication in this case?

	None
	More than 75%
	25-50%
	51-75%
	Less than 25%

peachmarten804

Please answer all

Why does a template-dependent DNA polymerase require a primer to initiate DNA synthesis?

	DNA polymerase require 5' phosphate to add a new nucleotide
	DNA polymerase require 3' hydroxyl group to add a new nucleotide

DNA polymerase require energy by hydrolizing the primers

Which of the following enzymes is template independent?

	Reverse transcriptase
	Klenow fragment

	DNA polymerase
	Terminal deoxynucleotidyl transferase

RNA polymerase

The natural function of a DNA ligase enzyme is:

	Joining together DNA sequences with compatible cohesive ends from two different organisms to yield a recombinant DNA molecule
	Sealing single stranded nicks in double stranded DNA molecules

	Joining together two blunt ended fragments of DNA within a cell
	Joining together two single stranded DNA molecules

The natural function of 3' -> 5' exonuclease activity of a DNA polymerase is to:

	Remove the 5' end of the DNA strand that is being copied
	Remove damaged nucleotide from the template strand

	Remove nucleotides from the end of DNA to generate blunt ends
	Remove incorrect nucleotides from the newly synthesized DNA

Which technique is used to resolve the different sizes of DNA fragments?

	Gene cloning
	PCR

	DNA sequencing
	Gel electrophoresis

Which of the following vectors does not contain bacterial origin of replication (oriC)?

	plasmid
	cosmid

	bacterial artificial chromosome
	lambda vector

Which process of bacteriophage is not used for gene cloning?

	Concatemer formation
	Lytic cycle

	Lysogenic cycle
	viral packaging

What vector would be best suited for creating a contig of bovine (cattle) chromosome 10?

	lambda vector
	Cosmid vector

	P1 vector
	Plasmid

E. coli takes up plasmid DNA by which of the following methods?

	Transduction
	Transformation

	Conjugation
	Transfection

The following times and temperatures are an example of the steps for PCR. You can use the Figure to help you answer the following questions.

Why is the first step is carried out at 94Â°C?

	To degrade the template
	To denature the template

	To activate the polymerase
	To facilitate the primer binding

What happens in the reaction when the temperature shifts to 55Â°C during cycling?

	the DNA polymerase will carry out DNA synthesis by extending the annealed primers
	the DNA polymerase will finish DNA synthesis

	the primers anneal to the single-stranded regions of the DNA
	the primers anneal to the double-stranded regions of the DNA

During cycling, what occurs when the temperature is at 72Â°C?

	the primers anneal to the double-stranded regions of the DNA
	the DNA polymerase will carry out DNA synthesis by extending the annealed primers

	the primers anneal to the single-stranded regions of the DNA
	the DNA polymerase will finish DNA synthesis

You designed a set of primers to amplify 1 kb DNA with polymerase chain reaction. By which cycle of polymerase chain reaction, we would expect to see the first double stranded DNA with expected size?

	The end of cycle 1
	The end of cycle 2

	The end of cycle3
	The end of cycle 4

1. You have the following prokaryotic DNA sequence: 5â²-GGACTAGACAATAGGGACCTAGAGATTCCGAAA-3â² 3â²-CCTGATCTGTTATCCCTGGATCTCTAAGGCTTT-5â² If the top strand is the template strand which sequence below is the correct mRNA transcript? A. 5â²-GGACUAGACAAUAGGGACCUAGAGAUUCCGAAAâ3â² B. 5'-UUUCGGAAUCUCUAGGUCCCUAUUGUCUAGUCC-3' C. 5'-CCUGAUCUGUUAUCCCUGGAUCUCUAAGGCUUU-3' D. None of the above are correct

2.Starting in the cytoplasm with a fully mature mRNA explain the process of translation initiation and then elongation and termination to me.

3.Prior to export from the nucleus, pre-mRNA must be processed. What specifically does this entail?

4.The biosynthetic pathway for the two amino acids E and H is shown schematically in the figure. You are able to show experimentally that E inhibits enzyme V, and H inhibits enzyme X. The most logical hypothesis regarding the regulation of Enzyme T would be that it is most likely subject to feedback inhibition by E alone. True or False?

5. A. kinase B. ligase C. polymerase D. oxido-reductase E. Phosphatase F. protease G. nuclease H. ATPase 1. Removes a phosphate group from a molecule 2. Hydrolyzes ATP 3. Hydrolyzes bonds between nucleotides 4. Adds phosphate groups to molecules 5. Catalyzes reactions in which one molecule is oxidized and another is reduced 6. Hydrolyzes peptide bonds 7. Joins two ends of DNA together 8. Catalyzes the synthesis of polymers such as RNA and DNA

6. Looking at the figure showing the change in reaction rate for two different enzymes that both utilize the same substrate, answer the following questions. a. What is the approximate Vmax for enzyme B ? b. What is the approximate Vmax for enzyme A ? c. What is the approximate Km for the enzyme which has a higher affinity for the substrate uM.

7. Answer the following question either yes or no (spell out the word, do not abbreviate as y or n). Looking at the following coupled reaction: Luciferase luciferin + O 2 ==========> oxyluciferin + light âG = + + Mg2+ ATP <=> AMP + PPi âG = - - - a. Based on your knowledge of free energy change and reactions would this coupled reaction occur ? b. Would changing the second step from ATP<=> AMP + PPi âG = - - - to ATP <=> ADP + Pi âG = - change your answer

7.Spontaneous reactions by definition will happen instantaneously. Why false?

8. Each of the following questions apply to the image of the channel protein. The purple sphere is a charged ion. Type your response to each question in the blank provided. a. Based you on what you know about cells and ions, what does the tri-lobed red and white molecule represent (although there aren't anywhere near enough of them shown) ? b. Since the question stated the purple sphere was a charged particle, using what you know about the tri-lobed red and white molecule what is the charge of the purples sphere c. On the extracellular space, on the right side of the purple sphere, there are two tri-lobed red and white molecules in close proximity to one another. The red of one appears to be attracted to the white of the other. What type of bond is this ? d. Based on your knowledge of proteins that interact with the membrane and the properties of the membrane, to what family of amino acids would the amino acid marked by A. most likely belong? ?

ZOL 341 Lecture Notes - Lecture 5: Consensus Sequence, Dnab Helicase, Origin Of Replication

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