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cell bio part 2 > Analyzing the Structure and Function of Genes > Flashcards

Analyzing the Structure and Function of Genes Flashcards

1
Q

What does a restriction enzyme do?

A

Cuts DNA strands at specific sites

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2
Q

Different enzymes recognize different sites and cut _________

A

At predictable locations

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3
Q

Restriction enzymes can produce what kinds of ends

A

Blunt or sticky ends

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4
Q

When restriction enzymes produce blunt and sticky ends, where are they

A

In DNA fragments

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5
Q

What the blunt and sticky ends in DNA fragments are useful for what

A

DNA cloning =

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6
Q

Who were the sole authors on the nobel prize paper on restriction enzymes

A

Daisy Roulland and Werber Arber

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7
Q

Who ended up getting the nobel prize on restriction enzymes

A

Arber, Nathans and Smith

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8
Q

What is electrophoresis

A

the separation of fragments based on size

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9
Q

What is used to separate fragments

A

Electrical currents

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10
Q

Negative charged of DNA is attracted to what

A

The positive electrode

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11
Q

Do smaller or larger fragments move through gel electrophores faster?

A

Smaller

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12
Q

Pieces of DNA can be “glued” together to produce what?

A

Recombinant DNA

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13
Q

Recombinant DNA is the basis for what?

A

GMO’s

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14
Q

Does recombinant DNA work for any DNA?

A

Yes any DNA from any cell

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15
Q

What is DNA cloning?

A

The transfer of recombinant DNA into an organism to replicate that DNA

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16
Q

What can DNA cloning express?

A

Human genes in easy to grow cells to study gene functions

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17
Q

What can DNA cloning express?

A

Human genes in easy to grow cells to study gene functions

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18
Q

An extension of DNA cloning is to create what?

A

A library of DNA cloning

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19
Q

What are restriction enzymes used for in DNA cloning?

A

To digest a chromosome

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20
Q

The DNA fragments are each cloned into what?

A

A plasmid

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21
Q

DNA cloning can also be done with RNA to study what?

A

Which genes are expressed in different conditions

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22
Q

What is a polymerase chain reaction (PCR)?

A

A powerful tool for amplifying large amounts of DNA for various reasons

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23
Q

PCR is basically ________

A

DNA replication in a tube

24
Q

PCR requires fewer what then for cellular DNA synthesis

A

Enzymes and proteins

25
What is only needed for PCR
DNA polymerase
26
A fragment of the DNA template is copied but what happens with the chromosomes
The whole chromosomes are not amplified
27
PCR is targeted what?
DNA replication
28
What are the three temperature settings that PCR requires?
Denaturation, annealing, extension
29
What is denaturation?
High heat breaks H bonds holding double strands together
30
What is annealing?
Cooler temperature for primer binding
31
What is extension?
Temperature that DNA polymerase works best to allow efficient and accurate synthesis
32
Uses for PCR testing?
Amplification for DNA cloning, Diagnostic testing, Forensic science
33
What are the two types of DNA sequencing commonly used for PCR testing?
Sanger sequencing and Next generation sequencing
34
What is Sanger sequencing?
Good for sequencing small pieces of DNA, commonly found and used in cellular, molecular and micro labs, and older but reliable technology
35
What is next generation sequencing?
Very good for sequencing large pieces of DNA and whole genomes, not as common due to cost, expertise, and application, new technology
36
How does sanger sequencing work?
Similar to PCR, but dNTPs are added, ddNTPs cause random chain termination during synthesis, and peices are separated and "end" NTP's at the end of the pieces are identified by color.
37
How does next generation sequencing work?
Highly parallel = more sequencing done at one time (in one reaction), Generates massive amounts of data, whole genomes can be sequenced in days to weeks, requires high power computer analysis skills.
38
NGS is like performing only short PCR sequencing runs, but instead you are doing what
Performing hundreds of thousands of them at once
39
NGS can quickly compare normal vs cancerous and even what type of cells
Metastatic
40
What does SDS-PAGE stand for?
Sodium dodecyl sulfate - polyacrylamide gel electrophoresis
41
What is ChIP
Chromatin immuno precipitation
42
What is EMSA
Electrophoretic mobility shift assay
43
What does FISH stand for
Fluorescent in Situ Hybridization
44
What can situ hybridization idenfity
can identify when genes are expressed in a cell or where genes occur on a chromosome
45
Uses what to base pair with nucleic acids in the cell
Fluorescently- tagged nucleic acids
46
What are reporter genes?
Genes that produce proteins with distinguishable traits are fused to promoters or regulatory regions of genes of interest
47
Gene deletion
Genes of interest can be deleted from the chromosome
48
What can you observe in gene deletion
Observe the phenotype of the organism to see how the mutation affects the phenotype.
49
RNA interference
use of RNA molecules to complementary bind specific mRNA in cells, which inhibits translation of that mRNA
50
What does RNAi create
creates double-stranded RNA, which is degraded by cells
51
What is RNAi similar too
Similar to gene deletion but less permanent
52
What else can RNAi do
Observe the phenotype of the cell to see what function or feature is lacking when the mRNA blocked from being translated
53
What is CRISPR
A mechanism to specifically edit the genome of an organism
54
What can CRISPR be used for
Can be used to remove genes and also can be used to add/replace genes
55
CRISPR is a natural system used by bacteria to fight what
viral infections
56
Adenoviral Vectors
Replace "sickening" genes with sgRNA and SpCas9
57
MOI (multiplicity of infection)
Ratio of virus particles to cells for infection

cell bio part 2 (9 decks)

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