CMMB 511 Lecture Notes - Lecture 5: Chloramphenicol, Transfection, Caat Box

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rosefrog223

28 Dec 2019

School

University of Calgary

Department

Cellular, Molecular and Microbial Biology

Course

CMMB 511

Professor

Lashitew Gedamu

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Document Summary

Transcription by rna polymerase 2 is enhanced by specialized gene activation sites. You can map presence of regulatory sequences using certain technologies. To do this, you introduce promoter proximal elements into a plasmid. The plasmid backbone has a promoter region until the cap site, then a goi that you want to understand how it is regulated. Take plasmid and transfect certain culture cells (eg. mouse cells, human cells). Sometimes gene may have protein factors of a certain type of cell, and it is in those specific cells the gene will be activated. The gene will be transcribed to five you mrna which can be translated into protein, but in the cultured cells you can measure the mrna synthesized using a hybridization assay. The mrna levels will indicate how a gene is being regulated (ie. lots of mrna means gene is active). You can mrna by different means such as northern blot,

Related Questions

Which type of RNA is unique to eukaryotes?

		mRNA
		rRNA
		tRNA
		snRNA
		Its a trick, all cells have all four types of RNA.

1 points

QUESTION 2

The region of DNA to which RNA polymerase binds to initiate transcription is the:

		origin
		promoter
		enhancer
		none of the above are correct

1 points

QUESTION 3

Holoenzyme:

		is RNA polymerase core enzyme.
		is sigma factor.
		is core plus sigma factor.
		none of the above are correct.

1 points

QUESTION 4

Which of these is formed FIRST when RNA polymerase binds to a promoter?

		The closed promoter complex
		The open promoter complex
		A rho-independent terminator
		none of the above are correct.

1 points

QUESTION 5

A hairpin loop followed by a run of Us:

		is the sequence formed in the sense strand of a gene using a rho-independent terminator.
		is the sequence recognized by the rho protein.
		is the structure formed in RNA that causes rho-independent termination.
		is formed immediately after the open promoter complex is formed.

1 points

QUESTION 6

RNA polymerase:

		reads the template strand of the DNA in the 3' to 5' direction.
		synthesizes the RNA in the 5' to 3' direction.
		can begin transcription without a primer.
		all of the above are correct.

1 points

QUESTION 7

Which of the following statements is FALSE?

		Bacteria like E. coli have only one type of sigma factor.
		RNA polymerase has no proofreading activity.
		The promoter of a gene is upstream of the RNA-coding region.
		Bacterial transcriptional terminators are copied into RNA to cause termination.

1 points

QUESTION 8

If the template strand of a gene has the sequence 5' ATGCGAT 3', then:

		the RNA produced will have the sequence 5' AUGCGAU 3'.
		the RNA produced will have the sequence 5' UAGCGUA 3'.
		the RNA produced will have the sequence 5' AUCGCAU 3'.
		None of the above are correct.

1 points

QUESTION 9

The sense strand:

		is the lagging strand
		is also the template strand.
		always goes left to right.
		none of the above are correct.

1 points

QUESTION 10

Which of these is transcribed but not translated?

		mRNA
		tRNA
		rRNA
		all of the above
		tRNA and rRNA are transcribed but not translated.

please help i am a bit confused

Which of the two major categories of the chemical reactions in photosynthesis is oxygen the INDIRECT product?

Glycolysis

Oxidative phosphorylation

Both light and dark

Light independent phase (LIP)

Light dependent phase (LDP)

You have a stomach flu characterized by vomiting. Which of the following systems is most responsible for the food in your stomach going the wrong way, e.g. vomiting? (Disregard this question and answer choice B)

Cardiac Muscle in the large intestine as discussed in class.

Smooth muscle in the gastrointestinal tract

Smooth muscle in the blood vessels

Skeletal muscle in the intestine

Proteins are made from which of the following smaller sub units?

Nucleotides

Amino acids that cannot enter the blood stream and go to cells

Monosaccharides

Fatty acids

Amino acids which can form new protein

Carbohydrates are digested in the gastrointestinal system into which of the following substances that is the smallest structure?

Polysaccharides that are produced by glucagon

Monosaccharides that are produced by insulin

Monosaccharides

Polysaccharide

Which of the following biomolecules has the longest digestion time? In other words, how long does it take for the biomolecule to be catabolized?

DNA

Lipid

Carbohydrate

Protein

You are sunburned after a long day at the beach. Which one is not an example of UV damage to your skin cells?

A mutation can occur in your ovaries

A mutation can occur in skin cell DNA that can lead to errors in transcription.

Errors can occur during DNA replication.

UV light can cause mutations in DNA of skins cells.

Thymine bases in your skin DNA bond with each other instead of across the double helix

This region, the ______________ (a) tells RNA polymerase where to bind to DNA while this region, the ______________ (b) tells RNA where to start transcription.

(a) promoter, (b) open reading frame

(a) start codon, (b) methionine

(a) promoter, (b) start site

(a) start site, (b) promoter

The genetic code is used to translate ______________ into ______________.

lipids, nucleotides

nucleotides,carbohydrates

nucleotides, lipids

nucleotides, amino acids

nucleosides, amino acids

Transcription of a gene has just completed. What are the next steps (in the correct order) to complete expression of that gene?

RNA polymerase makes a copy of DNA into mRNA.

mRNA is translated in the nucleus and ribosomal RNA moves into the cytoplasm.

Gene expression is completed after transcription.

mRNA moves into the cytoplasm and with tRNA they interact to form protein

Translation machinery moves into the nucleus and mRNA is translated into protein.

How is it that a mutation in DNA stored inside the nucleus of a cell can cause a disease affecting multiple organs?

This remains an unanswered question in science that scientists are still trying to figure out.

The mutated DNA is expressed in multiple cells through the process of transcription and translation, and the altered protein causes the disease wherever the DNA was expressed.

The mutated DNA is expressed in one cell and the altered protein travels to multiple organs to cause disease.

The mutated DNA is expressed and the mRNA takes the mutated information to other cells where the altered protein is made to cause disease in that organ.

The advantage of sickle cell anemia is which of the following?

It decreases the amount of oxygen in the blood cells so that mosquitoes carrying malaria will live

It decreases the amount of oxygen in the blood cells so that mosquitoe's blood form will die.

It allows the blood vessels to decrease the amount of blood going to the toes and feet.

There is none.

Which of the following is a purpose of cell division?

Cell reproduction

Growth

Repair

All of these

Use this list of gene elements to answer the following questions: (a) Promoter
(b) Exon
(c) Enhancer

(d) Intron
(e) 5â untranslated region

(f) 3â untranslated region

Problem 1. Which of the above gene elements are transcribed? You may choose more than one answer.

Problem 2. Which of the above gene elements are translated? You may choose more than one answer.

Problem 3. Which of the above gene elements are regulatory? You may choose more than one answer.

Problem 4. Which of the above gene elements are coding? You may choose more than one answer.

Problem 6. A mutation occurs in a gene with the following sequence: AAGCT. An example of a deletion mutation in this sequence would be AACT (loss of G). Using AAGCT as your normal sequence, give an example for the following types of mutations

(a) Insertion (b) Transition

Problem 7. A mutation occurs in the coding region of a gene with the following sequence AAGTGC. The translated sequence is Lysine (AAG), Cysteine (TGC). Give an example of following types of mutation using AAGTGC as your normal sequence. Provide the translated sequence caused by the mutation. (Hint: You will need a codon table to translate the sequence)

(a) Synonymous mutation
(b) Non-synonymous missense mutation

Problem 8. Patient 1 has a tumor causing mutation in a pro-growth gene. The mutated protein allows the cell to grow without receiving growth signals from outside the cell. Is the mutated gene an oncogene or a tumor suppressor? Why? Which hallmark is the gene associated with?

Problem 9. Patient 2 has a tumor which contains a loss of function mutation in the P53 gene. How would you determine if it is a somatic or germ line mutation?

Problem 10. Patient 3 has a tumor causing mutation in a gene called telomerase. The function of telomerase is to lengthen the ends of chromosomes. What hallmark is telomerase associated with? Is the mutation gain of function or loss of function? (Hint: to answer this question, you need to understand which hallmark telomere length is associated with)

CMMB 511 Lecture Notes - Lecture 5: Chloramphenicol, Transfection, Caat Box

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