Study Questions Set 1 Flashcards

1
Q

What information could you obtain from a genetic approach of studying mutants defective in a particular process?

A
  • the potential function and “importance” of the protein  identification/isolation/survival of mutants
  • how many genes are involved in a certain process
  • order of these genes (proteins) in the process by applying genetic approach (complementation analysis)
  • interaction between different genes (proteins) by using genetic approach (genetic suppression)
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

How would you define permissive conditions in respect to temperature sensitive mutants?

A

-permissive conditions are the temperatures at which the mutants can survive and grow

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

Define (or compare and contrast): a) gene expression; transcription; replication; translation; b) gene; allele

A
  • Gene Expression: the processes through which the information carried by a gene is converted into an observable product (either RNA or protein)
  • Transcription: the process of copying DNA to RNA by enzymatic processes
  • Replication: the process of copying a double stranded DNA molecule (DNA synthesis)
  • Translation: the process that converts an mRNA sequence into a chain of amino acids that form a protein
  • Gene: the functional unit of heredity; the entire DNA sequence necessary for the synthesis of a RNA molecule or functional polypeptide (it includes coding and regulatory regions)
  • Allele: different version of the same gene
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

Explain by using your own words the meaning/significance of gene expression.

A
  • gene expression is the conversion of a sequence of DNA into a final observable product (either RNA or protein)
  • it is important since all living things rely on proteins to function and survive  everything in/about the living cell/organism is about proteins
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

What are the roles of model organisms in molecular biology studies? Chose two model organisms and explain your reasoning.

A
-model organisms help us understand (at a basic molecular level) how something affects/alters cells and the organism in general; can be extrapolated to more complex organisms
E. coli:
- short reproductive period
-simple
-relatively cheap
-easy to maintain
-lots of data available
-4 million bp
-4000 different proteins
-divides every 20-60 minutes
-simple nutritional requirement
Drosophila:
-useful for genetic experiments
-developmental studies  body plan formation
-relatively small
-cheap
-easy to maintain
-lots of data available
-first to be extrapolated to higher mammals
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

What are three main functions of DNA? Explain the importance of each of them.

A

storage of information – sequence of bases codes for proteins, RNAs, regulatory signal

  • Preservation of information: replicates faithfully. semiconservative replication is basis of reproduction and cell growth
  • Variability of information - ability to mutate, basis of evolution and allelic variation
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

What is (are) the role(s) of phosphodiester bonds in DNA structure? What is (are) the role(s) of hydrogen bonds in DNA structure? What is (are) the role(s) of hydrophobic interactions in DNA structure?

A
  • phospho-diester bonds contain a negative charge  thus, the backbone contains a negative charge that repels two different strands from each other; rigid covalent bonds that are basis of structure
  • hydrogen bonds keep the two strands held together – complementary base pairing
  • the hydrophobic interaction helps stabilize the molecule since it forces the nitrogenous bases together and keeps the two negatively charged backbones apart
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

What noncovalent interactions are involved in maintaining the double-helical conformation of DNA

A
  • Van der Waals interactions: involved in the stacking interactions between bases
  • Hydrophobic interactions: the hydrophobic bases are kept on the ‘inside’ and the polar phosphate backbone is kept to the ‘outside’, to minimize the bases’ exposure to water
  • Ionic interactions: the DNA backbone is (-) charged, so salts (+) stabilize the helix by shielding it
  • Hydrogen bonding: the bond which is responsible for complementary base pairing
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

Describe Meselson-Stahl experiment and explain how it showed that DNA replication is semiconservative?

A

-E. coli cells were grown on media containing 15N for several generations  this made the DNA molecules denser than normal  cells were then grown for two generations on 14N  if the DNA got degraded, then the newly synthesized DNA would contain one hybrid band between the two nitrogen densities  since two different bands resulted, it confirmed that DNA replicates semi-conservatively  two strands of a parental DNA separate and each serves as a template for synthesis of a new daughter strand by complementary base pairing

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

What is meant by saying that a DNA strand has polarity? That two strands of DNA are antiparallel? That the strands are complementary to one another?

A
  • 5’ (phosphate) -> 3’ (hydroxyl)
  • at either end of the helix, the 5’ end of one strand will pair up with the 3’ end of another strand
  • the base pairs form hydrogen bonds with each other, such that A will always associate with T and C will always associate with G complementary bases
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

If a C content of a preparation of double-stranded DNA is 20%, what is the T content?

A
- Chargaff’s Rule: 
#purines = #pyrimidines
#A=#T
#C=#G
A+T =/= G+C (could be, but cannot be assumed)
So:
C=G so 20% C and 20% G = 40%
A+T must = 60%
If A=T then T content is 30%
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

What are the characteristics of B-DNA?

A
  • antiparallel
  • right-handed
  • long & narrow
  • 10 bp/turn
  • 0.34nm between bases
  • Pitch: 3.40nm
  • diameter: 2.0nm
  • tilt: 0 degrees
  • most of the in vivo DNA
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

What are the characteristics of A DNA?

A
  • antiparallel
  • right-handed
  • short & wide
  • 11 bp/turn
  • 0.23nm between bases
  • Pitch: 2.82nm
  • diameter: 2.3nm
  • tilt: 20 degrees
  • in low water content, can exist locally & temporarily in vivo, in dormant spores of Bacillus spp.
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

What are the characteristics of Z DNA?

A
  • antiparallel
  • left-handed
  • elongated & very narrow
  • 12 bp/turn
  • 0.38nm between bases
  • Pitch: 4.56nm
  • diameter: 1.8nm
  • tilt: 9 degrees
  • in test tubes with very high [salt]; in promoter regions of some genes: involved in regulation of gene expression
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

What are the characteristics of triple helical DNA?

A
  • formed when purines make one strand and pyrimidines the other (allows for third strand accommodations in the major grooves)
  • In introns of genes coding for cell membrane, phosphorylation, signal transduction, development, and morphogenesis proteins
  • Involved in regulation of gene expression
  • Used in gene therapy
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

How does high salt concentration influence denaturation kinetics of DNA? Explain your reasoning.

A
  • high salt concentrations (positive ions) impede denaturation, since these positive ions help to stabilize the DNA phosphodiester backbone
  • low salt concentrations reduce the stabilization and thus promotes denaturation of double stranded DNA
17
Q

What are the classes of DNA sequences in genomic DNA?

A
  • slow (unique): have lots of difficulty finding each other during renaturation (slow renaturation)
  • middle (moderately repetitive): moderate #, find each other with relative ease, need more time (middle renaturation)
  • fast (highly repetitive): short repeats of bp, no apparent function -> lots of them and they are able to find each other really easily (fast renaturation)
18
Q

What is Cot analysis?

A
  • Cot measures the rate of renaturation, which is a measure of the complexity of the DNA/genome
  • simple sequences renature more quickly than complex sequences
  • less complex sequences = repetitive sequences of nucleotides
    • complexity = # of nucleotides
  • more complex sequences = all unique nucleotides
    • complexity = # of unique nucleotides + # of nucleotides from ONE COPY of EACH repetitive sequence
19
Q

Who received a Nobel Prize for 3D DNA structure?

A

-James Watson, Francis Crick, and Maurice Wilkins (not Rosalind Franklin)

20
Q

If you had two solutions of DNA, one single-stranded and one double-stranded, with equivalent absorbance at 260 nm, how would the concentrations of DNA compare in these two solutions?

A
  • in single stranded DNA, the bases are unstacked and absorbance increases (hyperchromic effect)
  • in double stranded DNA, the bases are stacked and absorbance is reduced (hypochromic effect)
  • thus, to have equivalent absorbances, the concentration of double stranded DNA must be high, while the concentration of single stranded DNA must be low
21
Q

What is the difference between nucleoside and nucleotide? What does dNTP stand for?

A

Nucleoside: the sugar base and the nitrogenous base
Nucleotide: the sugar base, nitrogenous base, and the phosphate
dNTP: stands for deoxy-nucleoside triphosphate