BIOA01H3 Lecture 25: Module 3 Lecture 25.docx

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8 Feb 2015

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Overview of lecture 25: hershey/chase experiment: dna is hereditary material, dna structure and watson and crick model, meselson and stahl experiment: dna replicates semi conservatively, dna replication process, taq polymerase and the pcr, gel electrophoresis. If hereditary material is found in dna, then the hereditary material will contain radioactive phosphorus: bacteria infected with phage that had s 35 did not contain radioactive sulfur, bacteria infected with phage that had s 32 did contain radioactive phosphorus, progeny phage: bacteria infected with progeny phage containing 32 p were reactive. Gel electrophoresis processes: prepare agarose gel in between two electrodes, add buffer to cover the gels, load dna solutions, and marker dna, apply an electrical current, the dna fragments are negative so, they move to the positive side, shorter fragments move faster than longer ones, the gel is then stained and placed under special lighting to see.

Related Questions

Chapter 10

1.Outline the history of our knowledge on DNA up to Watson and Crick. What were the main contributions made by each researcherâs key experiment?

2.Explain the setup of the Hershey and Chase experiment, what would the results have been if protein was the genetic material?

3.Draw the structure of a DNA nucleotide, labeling each main component correctly. How does an RNA nucleotide differ?

4.If a section of double stranded DNA contains 19% Adenine, how much Thymine is present?

5.You are a researcher studying the genetic basis of heart attacks and have been working to determine the expression levels of different genes that might contribute to cancer formation. You obtain the DNA methylation status of five genes of interest (the data are shown in the table below). The plus (+) sign indicates the level of DNA methylation; more plus signs correlates with increased methylation levels.Based on this information which genes would you predict to have the highest rate of transcription?

Gene	Methylation levels
1	++
2	+++++
3	+++
4	++
5	+

What are the characteristics of the 3 main DNA forms?

Chapter 11

What are the different types of chromatin?

What are the structures and important roles for telomeres and centromeres?

What are the differences found between eukaryotic chromosomes and mitochondrial?

Chapter 12

Explain each of the different models of replication.

If you grow a culture of bacteria in media with radioactive nucleotides so that all DNA in the cells include radioactive nucleotides and then place the bacteria in new non radioactive media. After two rounds of replication what proportion of the DNA molecules will contain radioactivity?

Summarize the similarities and differences between rolling-circle replication, theta replication and linear eukaryotic replication.

What are the functions of the different DNA polymerases found in eukaryotic cells?

Draw a replication fork and include all key components and orientations. (Leading/lagging strands, DNA helicase, RNA primer and DNA gyrase)

What is the Holliday model of recombination and what are the necessary steps?

Chapter 13

What are the different types of RNA and what roles do they play?

Describe the properties and functions of each of the RNA polymerases and how they differ depending on the organism.

Describe in detail the process and mechanisms of transcription in both prokaryotes and eukaryotes.

Chapter 14

What are the primary purposes of each of the three post transcriptional modifications that occur in eukaryotic cells.

What is alternative splicing and what role does it play in the cell?

How is ribosomal RNA processed after transcription?

How do siRNA and miRNA work, describe/draw out the process in detail.

magine that the year is 1928 and the scientific world is in an uproar trying to determine what component of the cell isresponsible for genetic inheritance. Is it DNA, protein, perhaps even RNA? You are busy in your lab, trying to help solve this crucial mystery that still plagues (then) modern day biology. Your work focuses on a microorganism, a bacterium named Streptococcus pneumoniae, and its effects on mice. There are two strains of S. pneumoniae (which is responsible for pneumonia), the âRoughâ strain (R) and âSmoothâ strain (S). The strains are very similar except the S strain has a smooth polysaccharide coat that enables the microorganism to avoid the organismâs immune system and cause illness (and death in mice). The R strain has a rough coat which is detected and dealt with by the immune system (and as a result causes no death in mice). You decide to perform a series of experiments using the R and S strain of S. pneumoniae involving injections of these strains in mice. Your results are documented in the figure below: living S strain of S. pneumoniae mouse dies of infection living R strain of S. pneumoniae mouse lives mouse lives S strain heat-killed living R strain mouse dies of infection living, pathogenic S strain recovered S strain heat-killed From these results, you figure out that something from the heat killed S strain is transforming the R strain into the virulent strain. What is this âTransforming principleâ? Is it related to the cellular component that was passed from one generation to the next? As you know, this was the initial experiment performed by Dr. Frederick Griffith that eventually led to the discovery of the true cellular component that is the genetic inheritance molecule as well as the âtransforming principleâ (Hint: Itâs DNA!).

1. Several experiments were required to demonstrate how traits are inherited. Which scientist or team of scientists first demonstrated that cells contain some component that can be transferred to a new population of cells and permanently cause changes in the new cells?

a. Griffith b. Watson and Crick c. Avery, MacLeod, and McCarty d. Hershey and Chase

2. From your results, which one of the cellular components can be determined NOT to be the component responsible for genetic inheritance? Why?

a. Proteins because these macromolecules become denatured with heat. b. DNA because these macromolecules form a single helix in solution. c. RNA because it's double stranded 100% of the time d. Sugars because glycolysis happens.

3. Which one of the following experimental trials below are considered to be controls for this experiment? a. S Strain grown in 10% ethanol medium. b. Living S strain; Living R strain; Heat Killed S Strain c. Living R Strain with heat killed S strain; Living S Strain d. Heat Killed S strain; Living R strain; R strain grown in 10% ethanol medium

4. Which scientist or team of scientists obtained definitive results demonstrating that DNA is the genetic molecule? a. Griffith b. Watson c. Crick d. Hershey and Chase

5. Hershey and Chase used radiolabeled macromolecules to identify the material that contains heritable information. What radioactive material was used to track DNA during this experiment?

a. 3H b. 14C c. 35S d. 32P

6. DNA molecules, like proteins, consist of a single, long polymeric chain that is assembled from small monomeric subunits.

a. True b. False

7. The polarity of a DNA strand results from the polarity of the nucleotide subunits.

a. True b. False

8. There are five different nucleotides that become incorporates into a DNA strand.

a. True b. False

9. Hydrogen bonds between each nucleotide hold individual DNA strands together.

a. True b. False

brownpanda894

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%

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BIOA01H3 Lecture 25: Module 3 Lecture 25.docx

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