BIO 215 Lecture Notes - Lecture 8: Microbiology, Pasteurization, Polymerase Chain Reaction

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carminebee748

27 Jan 2017

School

StFX

Department

Biology

Course

BIO 215

Professor

Velautham Karunakaran

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Document Summary

1 to find out whether dna is expressed (antibody probe) Gene library: a collection of restriction fragments from a single genome. This is only useful if you can find one specific gene. If you have a gene expressing too much that you want to control, you would use this to control it. You need to shut off the mrna by using a complementary antisense strand adding it to the normal gene strand, to stop expression of the particular gene. This makes an rna hybrid, blocking transcription so replication of the dna is stopped. A virus infects a cell, you compromise the virus of infected cell and your body. All you need is one gene for piggy back vaccines. Replace part of the viral dna with foreign genes, and use this engineered virus to deliver code for antigenic determinants. The host responds by producing antibodies, becoming immune. produces antibody"s to fight it.

Related Questions

Multiple answers to each question might be possible!

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.

One step of the Hershey/Chase experiment involved blending the virus/cell mixture before centrifugation and probing the pellet for radioactivity. Why was the blending step necessary?

	To collect the bacteria at the bottom of the tube.
	To break open the bacteria to release the genome.
	To separate the bacteria from the bacteriophages.
	To be able to detect the radioactivity.

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.

Explain the 5â to 3â directionality of a DNA stand.

	It is due to the fact that the free 5â carbon is on one end and the free 3â carbon is on the other
	It is due to the fact that new nucleotide are added to the 5â carbon of the previous nucleotide
	It is due to the fact that there are 3 phosphate groups attached to the 5â carbon
	It is due to the fact the complementary strand is 3â to 5â
	More than one of the above explain the 5â to 3â directionality

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.

Why does DNA polymerase utilize an RNA primer?

	DNA polymerase is unable to initiate strand synthesis but RNA polymerase can
	DNA polymerase can only add a dNTP to an rNTP
	DNA synthesis proceeds in the 3â to 5â when initiating strand synthesis
	Chromosomal DNA contains interspersed RNA fragments
	The RNA primer increases stability of the newly synthesized strand

rosehippopotamus950

#1 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?

A) Using an RNA probe complementary to the region not removed by the truncation

B) Using an RNA probe complementary to the region removed by the truncation.

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

D) Using an DNA probe complementary to the region removed by the truncation

E) Using an antibody complementary to the region not removed by the truncation.

F) Using an antibody complementary to the region removed by the truncation.

#2To conduct the hybridization experiment, you are trying to decide between using a DNA or RNA probe. Which would be ideal to use and why?

A) As both are composed of nucleic acids, using either would result in identical results.

B) An RNA probe because RNA has uracil bases.

C) A DNA probe because it is more stable than RNA.

D) A DNA probe because RNA cannot bind to DNA.

#3Which of the following will lower the Tm of a given DNA strand?

A)Increasing the percentage of GC base pairs

B) Raising the pH of the solution from extremely acidic to neutral

C) Decreasing the buffer concentration from 50mM NaCl to 5mM NaCl.

#4 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?

A) No, because the RNA genome would not enter the bacteria upon infection

B) No, because while DNA and RNA nucleotides are similar, they are not identical

C) Yes, because DNA and RNA nucleotides are similar

D) Yes, because genome in any form (DNA, RNA, protein) would be labeled similarly.

#5 Griffith and Avery's transformation experiments allowed us to identify that DNA is our genetic information. Which of the following scenarios would result in bacterial cells that are capable of killing mice upon injection?

A) Heat killed non-virulent bacteria is added to a live virulent bacteria strain

B) Heat killed virulent bacteria is added to a heat killed non-virulent bacteria strain.

C) Heat killed mouse cells are added to a non-virulent bacteria.

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

A) Random DNA mutations generally won't affect RNA and protein function

B) It is faster to duplicate the genome when these are present.

C) The existence of introns can lead to multiple variations of proteins encoded by a single gene.

D) It is unlikely transposons would exist in the genome if there was too much protein coding DNA.

harlequineel21

The Case: While Elizabeth is reading the morning newspaper, she notices an ad for a free genetic screening for breast cancer at the clinic next week.The ad specifically invites women of Ashkenazi Jewish ancestry to participate.According to the newspaper ad, subjects will be tested to see whether they have mutations in the BRCA1 gene which would predispose them to breast cancer. Elizabeth, age 27, had heard about the discovery of the gene and about the mutation linked to Jewish women. Her paternal grandmother had been diagnosed with breast cancer at age 51 and died two years later, and Elizabeth worried that she may have inherited a variant that would put her at increased risk. She also worried about her mother, age 52 and apparently cancer-free so far, and her 7- year old daughter. Her daughter is not allowed to participate in the screening, but Elizabeth convinces her mother to go with her to get tested. For the screening, a small amount of blood is drawn. DNA is isolated from the blood, and part of the BRCA1 gene is amplified by PCR. Using a Southern Blot method, the digested DNA is run on a gel, transferred to a neutral membrane. Once on the membrane, the samples are hybridized with specific probes corresponding to deleterous mutations known to be linked to increased breast cancer susceptibility, plus probes for wild type regions of BRCA1. The probe will only bind to the DNA if that mutation is present. Control DNA samples, known to have a specific mutation, are included in the run to verify probe . Probe binding is visualized by labels attached to the probe, and will depend on the lab running the analysis. Probes: 185delAG (AG deletion in exon 2) Normal 185 (no mutation at this site) 4184delTCAA (TCAA deletion in exon 11) Normal 4184 5382insC (C insertion in exon 13) Normal 5382 DNA samples: Elizabeth Mother Unrelated woman Control 185delAG Control 4184delTCAA Control 5382insC Control wild type BRCA1 (no mutations) If either Elizabeth or her mother test positive, is a prophylactic double mastectomy appropriate? (2 marks) What other issues does this type of testing raise, and how should these issues be addressed? (2 marks) What does it mean if a result came back as a Variant of Uncertain Clinical Significance? Provide a peer-reviewed journal article as a reference for this question. (1 mark answer, 1 mark reference) Does the lack of any of these mutations mean that the women will not get breast cancer? (2 marks) Who should have access to the test results? (2 marks) Does the daughter have the right to know the results? She is only seven now, but what about when she is 16? (3 marks) Should this type of screening be mandatory? (3 marks)

BIO 215 Lecture Notes - Lecture 8: Microbiology, Pasteurization, Polymerase Chain Reaction

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