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Biomedical Lab > Lab 8 > Flashcards

Lab 8 Flashcards

1
Q

Proteins are made of ___ linked by ____

A

Amino acids linked by peptide bonds

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2
Q

Small amino acid chains are called…..

A

Peptides

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3
Q

Proteins are formed by ______ or _______

A

Lengthy single amino acid chains or several polypeptide chains

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4
Q

Peptides are proteins carry ____ groups on their side chains

A

Charged

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5
Q

What are the two charged groups on peptide and protein side chains?

A

COO- NH3+

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6
Q

Functional proteins have a _____ dimensional structure due to which 4 bonds?

A

3 dimensional
Hydrophobic, ionic, hydrogen, and disulfide bonds

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7
Q

Most proteins that deal with light use _____ to capture and release ______

A

Exotic molecules to capture and release photons

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8
Q

____ uses a special sequence of 3 amino acids:

A

GFP
Serine-Tyrosine-Glycine

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9
Q

In the presence of oxygen, what happens to these 3 amino acids?

A

They oxidize to form a fluorescent chromophore in the center of GFP

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10
Q

What happens when the GFP structure is changed and the chromophore is disrupted?

A

The fluorescence is lost

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11
Q

GFP absorbs ___and ____ light and emits ____ light

A

Absorbs blue and ultraviolet light and emits green light

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12
Q

Gene expression refers to…….

A

The entire process from transcription to protein synthesis

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13
Q

Genes essential for survival are generally expressed _______ (_____)

A

Continuously (constitutive)

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14
Q

The expression of most genes is……..

A

Regulated and controlled by other molecules

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15
Q

Gene regulation is a process that…..

A

Allows mRNA and protein synthesis only when and where the encoded protein is required

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16
Q

Give 2 specific examples of gene regulation in the body

A

-myosin in muscles for contracting
-amylase in salivary glands for carbohydrate digestion

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17
Q

One method of regulating gene expression is to….

A

Control the synthesis of mRNA (transcription step)

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18
Q

What is a promoter?

A

A DNA sequence upstream to the coding region of a gene to which RNA polymerase binds to initiate transcription

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19
Q

Each gene or set of genes requires its own unique ______

A

Promoter

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20
Q

When a gene is regulated, how is the function of RNA polymerase affected?

A

RNA polymerase cannot initiate transcription unless the regulatory molecules are present or when the inhibitor at the promoter region is released

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21
Q

What is responsible for differentiating stem cells to functional specific cells?

A

Regulation

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22
Q

How are stem cells differentiated into muscle cells??

A

In a stem cell, the protein actin is not present. As a result, the stem cells do not have the muscle phenotype. To become muscle cells, stem cells must express actin. Regulatory molecules are required to initiate actin transcription and differentiate stem cells to muscle cells

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23
Q

The regulatory molecule can either bind to the _____ or the ______ or the _____

A

Promoter or to RNA polymerase or to the inhibitor (allows transcription to happen)

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24
Q

in pGLO transformed bacteria, what 2 proteins are required to initiate transcription of the GFP gene?

A

L arabinose and the Ara C protein

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25
Gel electrophoresis is a procedure used to……
Separate biomolecules such as DNA, RNA, or proteins using a gel matrix agarose or polyacrylamide
26
In gel electrophoresis, separation depends on….
The properties of the gel as well as the molecules to be separated
27
The gel matrix contains….
Uniformly sized pores through which molecules can pass through
28
The size of the pores in a gel matrix depends on…..
The gel material
29
What determines the size of the pores?
Gel concentration
30
Polyacrylamide is generally used to separate ___ which are generally much ____ than DNA and RNA
Proteins which are generally much smaller than DNA and RNA
31
What is the MOST IMPORTANT factor for migrating through the gel matrix?
Size of the molecules
32
Which migrates more slowly - larger molecules or smaller molecules?
Larger molecules
33
The protein sample is mixed with ___ and loaded in _____
Mixed with a sample loading buffer and loaded in wells
34
The loading buffer is a concentrate containing __ and ___
A tracking dye and glycerol
35
The colored (blue) tracking die has a very low….
Molecular weight
36
What is the purpose of the glycerol?
Increases the density of the sample to make it fall easily into the wells and prevent it from floating away
37
What is used to determine the size of the proteins in the samples?
A protein molecular marker of known sizes
38
What are the 2 driving forces of gel electrophoresis?
Electric current Charge
39
Why is electrical current imperative in terms of gel electrophoresis?
Molecules require electrical current to migrate through the gel pores
40
CHARGE When a current is applied, positive molecules (cations) will move to the ____ (____ electrode) while negative molecules (anions) will move to the _____ electrode (____)
Cations will move to the cathode (negative electrode) Anions will move to the positive electrode (anode)
41
Molecules of the same size but different ___ will differ in migration
Charges
42
To separate molecules by size only, all the molecules must have a similar ______
Charge
43
Why is water not used for preparing gels or used as a running solution ?
It is not a good conductor of electricity
44
Running buffer has the _____ which ensure the conductance of electricity through the gel
Electrolytes
45
What are the 3 major techniques used to analyze and purify proteins???
-SDS page electrophoresis -centrifugation -chromatography
46
In SDS page electrophoresis, the matrix is made up of….
Acrylamide polymer
47
By controlling the percentage (____ to _____) of acrylamide polymer……..
3%-30% Precise pore sizes can be obtained to separate proteins form 5kDa-2000kDa
48
Laemmli buffer is designed to…..
Dissociate proteins into individual polypeptide subunits
49
What does SDS stand for in SDS page electrophoresis?
Sodium dodecyl sulfate
50
SDS is an…..
Ionic detergent which denatures protein complexes by wrapping around the polypeptide backbone
51
What does DTT stand for?
Dithiothreitol
52
DTT is a….
Thiol reagent that cleaves disulfide bonds
53
What happens when a protein sample is heated in presence of DTT and excess of SDS?
Proteins are FULLY dissociated into their subunits
54
____ provides the force for migration in SDS page electrophoresis
Electric current
55
The gel is run in a running buffer containing ___ and ____
Electrolytes and SDS(to mask charges)
56
Once the individual proteins are separated, the gel is stained with….
Coomassie blue ( GelCode Blue) to visualize the protein bands
57
Name the 5 components of the Lammeli sample buffer with DTT
SDS Bromophenol blue Glycerol Tris pH 8 DTT
58
The total protein is assayed using….
Reddish brown Coomassie blue dye
59
_______ amino acid residues in the protein interact with Coomassie blue dye
Nonpolar
60
The protein-dye complex causes a…..
Shift in the dye absorption maximum, from 465nm to 595nm to give a blue color
61
The intensity of the blue color is proportional to…
The protein concentration within a short linear range
62
The Bradford reagent gives….
Linear response from 1-140 micrograms
63
____ is commonly used for generating the standard curve
BSA (Bovine Serine Albumin)
64
How are the protein bands observed?
They’re stained with a dye (in this lab = GelCode blue)
65
GelCode blue stain is a proprietary ___ stain
COOMASSIE BLUE BASED STAIN
66
The GelCode blue also stains the…
Gel

Biomedical Lab (10 decks)

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