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Molecular Biology > Final Exam > Flashcards

Final Exam Flashcards

1
Q

What propagates bio. molecules, observes a physiological process, source material, etc. ?

A

Cell culture

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2
Q

What manipulates DNA?

A

DNA cloning

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3
Q

What obtains a sequence of monomers in a biomolecule?

A

DNA sequencing

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4
Q

What separates polypeptides by size?

A

SDS-PAGE

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5
Q

What Identifies a biomolecule by comparing it to sequences stored in a data base?

A

BLAST

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6
Q

What visualizes a specific protein in a cell-free extract?

A

Western blot

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7
Q

What uses a conjugated antibody to identify a protein?

A

Immunostaining

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8
Q

What makes millions of copies of a DNA sequence?

A

PCR

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9
Q

What identifies a specific protein in a bodily fluid such as urine or blood?

A

ELISA

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10
Q

What identifies a specific protein in intact cells using a conjugated antibody?

A

Immunocytochemistry

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11
Q

What identifies a specific protein in tissue sample using a conjugated antibody?

A

Immunohistochemistry

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12
Q

What protein is generated by a challenge to the immune system; binds to a specific biomolecule?

A

Antibody

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13
Q

Define transformation and explain why the mouse did not die when injected with the cell-free extracts of the pathogenic strain, but did die when injected with the pathogenic cell-free extract and the live harmless strain.

A

Transformation in when a cell picks up genetic info from another cell and incorporates it into its own genome. The mouse did not die because the cell contents of the pathogenic strain do not cause the harm. The harmless strain picks up the genetic info of the pathogenic strain and expresses it pathogenic characteristics.

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14
Q

How did Avery’s approach indirectly identify the “transforming” molecule as nucleic acids and not proteins?

A

Because when DNAse was mixed with the cell-free extract of the pathogenic strain and the harmless strain, the mouse did not die even though it was supposed to.

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15
Q

How did Hershey and Chase’s approach directly identify nucleic acids and not protein as the “transforming” molecule?

A

They determined that DNA is what enters the cell rather than protein. They did this by using different bacteriophages, one with a radioactive protein coat, and the other with radioactive DNA. Using a centrifuge they determined that the protein stayed outside of the cell and the DNA entered the cell.

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16
Q

Why is was the discover of T. aquaticus important for PCR?

A

Because its polymerases can function at high temperatures, while other polymerases cannot.

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17
Q

What are Messelson and Stahl known for?

A

Proving that DNA replication was semiconservative using isotopes. (Bacterias growth in media with heavy isotopes, then moved to media with lighter isotopes. Change in banding occurred.)

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18
Q

What are Sanger and Mullis known for?

A

PCR and DNA sequencing

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19
Q

What is Zamecnik known for?

A

protein synthesis in vitro

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20
Q

What is Hurwitz known for?

A

RNA synthesis in vitro

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21
Q

What is Nirenberg known for?

A

Cracking the genetic code

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22
Q

What is Kornberg known for?

A

DNA synthesis in vitro

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23
Q

What is Avery known for?

A

identified (indirectly) the “transforming principle” as a nucleic acid using the enzymes DNAse, RNAse, and proteases. (still used bacteria and mice)

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24
Q

What is Griffith known for?

A

transforming a harmless strain of Streptococcus into a pathogenic strain….but he didn’t show which biological molecule was responsible

25
What are Hershey and Chase known for?
identified (directly) the transforming principle as a nucleic acid using unstable isotopes, a blender (to knock bacteriophages off of the bacteria), and a centrifuge.
26
Which level of protein structure are affected by a denaturant?
2,3,4
27
What bonds are affected by a reducing agent?
disulfide bonds
28
Looking a western blot gel, where do heavy and light bands appear?
Heavy bands are at the top on the gel. Light bands are at the bottom on the gel.
29
What are primary cells and cell lines?
Primary cells are cells grown in media that come straight form the source. They respond to signals appropriately and have a finite number of replications. Cell lines are cells that have been kept alive after a lot of replication cycles. They do not respond to signals appropriately.
30
Why is the info in the major groove of DNA more informative than that in the minor groove?
Because it contains more observable patterns in the major groove than in the minor groove. The major groove also contains a larger space than the minor groove.
31
How did the MICA experiment determine the number of nucleotide residues that made up one complete turn of the double helix?
DNA was laid down on salt, which formed bonds with one side of the DNA helix. DNAse's were then added and degraded the other side of the DNA helix. The DNA was then recovered and more on a gel. The bands represented one nucleotide.
32
How does a high salt buffer change the melting temp (Tm) of a DNA molecule?
It stabilizes the negatively charged DNA and lowers the repulsion between the two strands.
33
Describe how the Tm is determined experimentally.
Tm is determined by when half of the DNA is double stranded and the other half is single stranded. This is done by taking the absorbance of the DNA as temp increases.
34
What is in Reagent I. of alkaline lysis?
CDTA- binds to divalent cations, ex. Mg2+ (stops DNAse) Lysozyme- weakens bacterial cell walls Tris- buffering agent (pH=8) Glucose- keeps solution at an isotonic state
35
What is in Reagent II. of alkaline lysis?
SDS- solubilizes phospholipids high [NaOH]- denatures proteins and all DNA
36
What is in Reagent III. of alkaline lysis?
Potassium acetate- neutralizes solution that leads to the separation on plasmid DNA from everything else
37
What is alkaline lysis?
A method to extract pure plasmid DNA from bacteria
38
What is the whole purpose of reagent I. of alkaline lysis?
Used to resuspend bacterial pellet
39
What is the whole purpose of reagent II. of alkaline lysis?
Denatures stuff and lyses the cells
40
What is the whole purpose of reagent III. of alkaline lysis?
separating everything from plasmid DNA
41
What are the axis labels on a flowcytometry graph?
y-axis= SSC (side scatter)- measures cell complexity x-axis= FSC (forward scatter)- measures cell size
42
Components of typical eukaryotic gene's and their function
Promotors- DNA sequence needed to allow a polymerase transcribe a DNA sequence Intron- region of DNA that do not code for proteins Exons- Regions of DNA that code for proteins Ori site- site on DNA where replication begins Terminator- Sequence that stops transcription
43
What are components of a recombinant plasmid?
Promoter terminator Target sequence Restriction enzyme sites One ORI site Antibiotic resistance gene
44
What does DNA ligase do?
"glue's" two DNA strands together
45
What does DNA polymerase do?
Used to replicate DNA to DNA
46
What does RNAse H do?
Used to remove primers
47
What does an exonuclease do?
Used to cut at a specific sequence which opens up the plasmid
48
What does reverse transcriptase do?
Used to convert RNA to DNA
49
Why is gene density higher in prokaryotes than eukaryotes?
Because prokaryotes contains a higher proportion of genes that encode for proteins compared to eukaryotes.
50
What does DNA acetylation do?
removes positive charge of the lysine on histones
51
What does DNA methylation do?
Enhances or inhibits proteins attaching to DNA
52
What does phosphorylation of DNA do?
adds a negative charge to serine on histones
53
Explain how transcription is initiated in prokaryotes
Prokaryote RNA polymerases contain to subunits. The sigma/initiation factor and the core RNA polymerase. The sigma factor binds to the recognition sequence on the promoter then recruits the core subunit.
54
Explain how transcription is initiated in eukaryotes
A transcription factor called TBP (TATA-bind protein) binds to the TATA box (a regulatory sequence). Then transcription begins
55
Describe the function of the two components of the prokaryotic RNA polymerase.
Sigma factor- Recognizes and binds to recognition sites in the promoter region and recruits the core polymerase. Core polymerase- Contains the catalytic sites and builds the mRNA strand.
56
Explain the composition of the open reading frame.
The open reading frame a sequence on the mature mRNA that is made out of codons that are flanked by UTR's (untranslated regions)
57
Explain how the ribosome gets recruited at the start site of translation in prokaryotic mRNAs.
Ribosomes bind to the shine dalgarno sequence and move along the mRNA until they get to the start codon
58
Explain how the ribosome gets recruited at the start site of translation in eukaryotic mRNAs.
Ribosomes binds to the 5' cap and moves down the mRNA until it meets the Kozak sequence which is upstream on the start codon
59
Explain how puromycin inhibits translation.
It is a chain terminator that get into the P site of a ribosome and causes premature termination of nascent polypeptide chains

Molecular Biology (3 decks)

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