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Chapter 2 Outline

1. Griffith's experiments with bacteria showed that heat-killed virulent cells could transform avirulent cells, indicating a genetic basis. Further experiments by Avery, MacLeod and McCarty found that the transforming agent was DNA. 2. Hershey and Chase used radioactive labeling to show that DNA, not protein, was injected by bacteriophages into bacterial cells during infection. 3. DNA is made up of nucleotides, which contain a nitrogenous base, a sugar (deoxyribose in DNA), and a phosphate group. Nucleotides are joined by phosphodiester bonds between the phosphate of one and the sugar of another.

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26 views

Chapter 2 Outline

1. Griffith's experiments with bacteria showed that heat-killed virulent cells could transform avirulent cells, indicating a genetic basis. Further experiments by Avery, MacLeod and McCarty found that the transforming agent was DNA. 2. Hershey and Chase used radioactive labeling to show that DNA, not protein, was injected by bacteriophages into bacterial cells during infection. 3. DNA is made up of nucleotides, which contain a nitrogenous base, a sugar (deoxyribose in DNA), and a phosphate group. Nucleotides are joined by phosphodiester bonds between the phosphate of one and the sugar of another.

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Brian Shee
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Chapter 2: The Molecular Nature of Genes

1. F. Griffith: Laid foundation for DNA identification as genetic material w/ transformation in


bacteria s. pneumonia
a. Heat killed virulent cells were able to transform avirulent cells to virulent
2. Avery, MacLead, McCarty used similar transformation method as Griffith to find transforming
agent
1. Removed protein (destroy protein)  still transformed
2. Trypsin & chymotrypsin digestion  still transformed
3. Tried ribonuclease to degrade RNA  still transformed
4. DNAse  destroyed transformation
i. DNA = transformation agent

3. Hershey & Chase experiment w/ T2 bacteriophage infection on e. coli


a. During infection DNA was injected, very little protein was injected
b. Used radioactive labels phosphorous 32 for DNA & sulfur 35 for protein
c. Labeled phages attached their tails to bacteria & injected their genes into their host
4. Chemical Nature of Poly nucleotides
a. DNA  nitrogenous base, phosphoric acid & sugar deoxyribose
b. RNA  nitrogenous base, phosphoric acid & sugar ribose
Thymine is only
one w/ a methyl
group

5. Nucleosides: joined units f bases & sugars from DNA or RNA


6. Ribose (RNA) vs. 2-deoxyribose (DNA)

7. Nucleotides: nucleosides with a phosphate group


Phosphodiester Bonds: Joins nucleotides together in
DNA or RNA via phosphoric acid linked to 2 sugars.

One @ the 5’ sugar, other 3’ sugar


8. DNA Structure
a. Chargaff’s studies found: # of purines =# of pyrimidine(A to T, G to C) GC content is
variable among diff species
i. Known as Chargaff’s rules
9. Franklin’s x-ray suggests DNA is a helix
a. Watson suggested DNA is a double helix w/ sugar phosphate backbones on the outside &
its bases on the inside
10. Spacing btwn base pairs is 3.32A, a turn is 33.2A (10.4bp per turn)
a. Antiparallel, semiconservative replication
11. Genes made of RNA
a. Certain viruses are made of RNA genes instead of DNA
12. Physical Chemistry of Nucleic Acids
a. B form: Proposed by Watson & Crick, represents sodium salt of DNA in a fiber at 92%
humidity (?). This form is the most abundant DNA form
i. The "salt" of DNA is the form in which some of the hydrogen ions have
disassociated from the phosphate group. The salt then has a net negative charge,
specifically at the oxygens of the phosphates
ii. Higher GC content = higher salt content
b. A form: if humidity is decreased to 75%, A form is assumed
i. Fatter & shorter, 10.7bp per turn, each turn is 24.6A
ii. Base pairs tilted away from the horizontal
iii. Hybrid polynucleotide of 1 DNA & 1 RNA
1. As well as dsRNA
iv. Both A & B are right hand twists
c. Z form: dsDNA w/ alternating purine and pyrimidine –GCGCGC-

-CGCGCG-

i. Can exist in an extended left handed helical form


ii. Longer & skinnier than B DNA
1. Living cells contain a small portion of Z DNA
2. Activation of at least one gen requires that a regulatory sequence switch
to the Z DNA form
13. DNA denaturation/DNA melting: When 2 strands come apart either by heating or other means
a. Tm: melting temperature, temp at which DNA strands are half denatured
i. Higher GC content, higher Tm
1. b/c GC forms 3 hydrogen bonds, AT forms only 2 H-bonds
b. Hyperchromic shift: increase in a DNA solutions’ absorbance of 260nm light upon
denaturation
c. Other ways to denature DNA
i. Heating
ii. Organic solvents i.e. Dimethyl sulfide, formamide
iii. High pH
iv. Disrupting H-bonds
v. Lowering salt conc of DNA removes the ions that shield the (-) charges
1. Mg2+ etc
d. Ways to anneal/renature:
i. Temperature (25oC below its Tm)
ii. DNA concentration: The higher the conc in solution, the more likely they will
encounter each other and anneal
iii. Renaturation time: the longer the time allowed for annealing, the more will occur
14. 660- approximate molecular weight of one avg nucleotide pair (multiply by bp to get molecular
weight)
a. Small DNA can be estimated by electron microscopy
i. Can also reveal if DNA is circular or linear, or supercoiled
15. AA avg 110 molecular weight, aprox 364 AA per protein
16. C value: DNA content per haploid cell
a. C-value paradox: C-value of a species is not necessarily related to the genetic complexity
of the species i.e. human vs. frogs
i. Probably explained by # of extra noncoding DNA

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