CEDB20003 Lecture Notes - Lecture 8: Wild Type, Tumor Suppressor Gene, P53

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greymarten981

13 Aug 2018

School

University of Melbourne

Department

Cell Biology

Course

CEDB20003

Professor

Jenny Gunnerson

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Document Summary

We can analyze the output of transcription (rna) or translation (protein). The methodology we employ will depend on what we wish to know: (what cells in a tissue are producing the mrna?) We can look at the output of transcription, rna. A lot of the tests are destructive, killing the cell; but what if we want to study it while it is. The amount of translation so the amount of protein found in the cell alive. Use a radioactive probe which is antisense to the mrna that you want to study so that they bind together. The hybridization allows you to detect where the mrna is by measuring the radioactivity. Quantitatively measure how much rna is within that tissue. How are we measuring the one specific mrna sequence: First we run it through a gel, passing an electro current so that mrna will migrate towards the positive end of the gel.

Related Questions

1. A study was conduced to identify a liver cell gene (zed) that has a length of 3000 bp. A southern blot was prepared using DNA from the liver cell and a probe made to region 1600-2000 of zed. Analysis of the blot revealed that there are two spots present, one at 3000 and another at 2000. A plausible explanation for this observation is

A. there is an identical copy of this gene on another chromosome.

B. the gene was cut into two equal portions by a restriction enzyme.

C. there is another gene with sequence similar to gene zed

D. both: there is an identical copy of this gene on another chromosome and there is another gene with sequence similar to gene zed

E. no explanation can be given using the available data

2. Protein P is synthesized in relatively high amounts in human pancreas. This protein has been isolated and purified, but its amino acid sequence has not been determined. We wish to clone the gene for protein P.

(a) How can a probe be prepared to identify the gene for protein P?

(b) If we have prepared a radioactive messenger RNA as our probe in part(a), how could we verify that it is the mRNA for protein P?

(c) If we wish to make a gene library from the DNA of human pancreatic cells, we proceed by digesting the DNA with the restriction enzyme BamHI and then separating the fragments by agarose gel electrophoresis. The fragments are transfered (by southern blotting) onto a nitrocellulose filter and flooded with the radioactive probe. Following incubation, the filter is washed to remove any probe that has not specifically bound to one or more DNA fragments. Autoradiography of the filter reveals two bands of size 300 and 700 bp. When experiment is repeated using restraction enzyme PstI, only one band of 1000 bp appears. If it is already known that pancreatic cells contain a single copy of the gene of protein P, which of these enzymes should be used to construct our gene library?

violethornet799

1. What reaction is catalyzed by reverse transcriptase and used in the laboratory to generate cDNA?

a. transcription of RNA to DNA

b. transcription of DNA to RNA

c. conversion of ribonucleotides into deoxyribonucleotides

d. translation of RNA to DNA

e. conversion of a ribonucleotide to a DNA double helix

2. In eukaryotic cells, _________determine the amount of mRNA arising from a particular gene.

a. transcriptional-level controls

b. RNA processing-level controls

c. translational-level controls

d. replication-level controls

e. b and c

3. In eukaryotic cells, _________determine how a primary transcript is converted to mature mRNA

a. transcriptional-level controls

b. RNA processing-level controls

c. translational-level controls

d. replication-level controls

e. a and d

4. In eukaryotic cells, _________determine when a particular mRNA is actually used in protein synthesis.

a. transcriptional-level controls

b. RNA processing-level controls

c. translational-level controls

d. replication-level controls

e. None of the above because protein synthesis takes place immediately

5. Under what circumstances do cells selectively activate and suppress their genes?

a. at different stages of embryonic development

b. in different tissues

c. exposure to different types of stimuli

d. No such circumstances exist in natureâcells express all their genes throughout life.

e. a, b and c

6. Which statement about the control of gene expression is FALSE?

a. A single gene may be controlled by different transcription factors.

b. A single transcription factor may be necessary for the expression of different genes.

c. Each cell type has a characteristic pattern of gene transcription, which is determined by the particular complement of transcription factors contained in that cell.

d. Expression of protein genes is primarily controlled in nucleoli

e. Control of gene transcription is complex and influenced by various circumstances.

7. What is thought to maintain X chromosome inactivation in females?

a. DNA acetylation

b. DNA methylation

c. RNA inactivation

d. Histone methylation

e. None of the above because both X chromosomes are active in females and males.

crimsonplatypus494

1. To understand the structure of DNA (deoxyribonucleic acid) we need to know and understand how the bases in DNA are composed. Please state Chargaffâs rules (there are two).

2. Explain why replication of the DNA is termed semiconservative.

3. Explain what transcription is and what translation is. Describe the three major steps in DNA replication.

1. Describe the three major classes of RNA (ribonucleic acid); what is the function of each class?

2. Describe the location of ribosomes and what their function is.

1. Discuss (to the best of your knowledge and understanding) what is a GFP (Green Fluorescent Protein) and how researchers might use a GFP-labeled protein to study cancer?

2. How would you test a hypothesis that a genetic condition, like cancer, is due to an error (or mistake) in transcription and translation?

CEDB20003 Lecture Notes - Lecture 8: Wild Type, Tumor Suppressor Gene, P53

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