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See Fig 33.5 Major pyrimidines and purines Tautomers of adenine and cytosine
Amino versus Imino
5 6
1
Tautomers of guanine, thymine and uracil
Lactam versus Lactim Ribose and Deoxyribose
Predominant forms
See 33.3 Figure
RNA DNA
7 8
Nucleosides
9 10
2
Structures of the deoxyribonucleoside-5’-
monophosphates
See Fig 33.7
13 14
(continued)
***
15 16
3
Backbone of
Nucleic Acids
Story of DNA as Genetic Material
Discovery of the structure of double
Fig 33.4 stranded DNA, 1953
James Watson, Francis Crick, Rosalind Franklin,
Maurice Wilkins
20
S = capsular (1)
polysaccharide Evidence for
= death transforming
principle!
R= no
capsule =
live Evidence that
(2) DNA is the
Isolation &
characterization
genetic
of the
transforming material in
principle proved
cells!!
the chemical
make-up of the (3)
genetic material
Both set of phage
were infective
(4)
21
4
see Fig 33.12
BCMB 3100 - Nucleic Acids - Chapter 33
•Discovery of DNA
Adjacent •Nucleotides, nucleosides & bases
nucleotides
can •Polynucleotides
hydrogen •DNA as genetic material
bond to
•Structure of double-stranded DNA
each other
•Chromatin
•RNA
•Nucleases
25 26
27 28
• Structure of B-DNA
• Sugar phosphate backbone
outside
• Stacking creates two
Complement unequal grooves (major
ary base and minor)
pairing and • Hydrophobic attraction
stacking in between the bases
DNA • Van der Waals contact
between bases
• H-bonds between bases
• Electrostatic repulsion
between phosphates
inhibited by cations (Mg++)
29 30
5
Double helix
Fig 33.13 emphasizing the charge
on the phosphate
groups
Fig 33.11
6
Fig 33.20
38
Minor groove: 6Å wide; 7.5Å deep Human 240 x 106 bp 3200 x 106 bp
Fig 33.19 (46 chromosomes)
7
BCMB 3100 - Nucleic Acids - Chapter 33 Stem-loop structures in RNA
8
Structure of supercoiled DNA. Circular B-DNA has 10.4
bases/turn of helix. If DNA is underwound (or overwound), it Human topoisomerase I bound to DNA
is supercoiled to restore 10.4 bases/turn. Supercoiling is done
by topoisomerases.
• Topoisomerases can
Supercoiled (underwound)
Relaxed; [supercoils are caused by underwinding or overwinding] add or remove
supercoils in DNA
(10.4 base pairs per turn
of the double helix)
Fig. 33.24
Electron micrograph of chromatin 53
9
Histone octamer Nucleosome core particle Fig. 33.26
Nucleosome
Nucleosome gives
10-fold packing
54 bp
146 bp
Fig. 33.25
57
59
10
Representation of the compaction of DNA Final chromosome is 1/8000 of length of B-DNA.
into a eukaryotic chromosome
This allows DNA to be packaged into cells. For
example, the largest human chromosome is 2.4 x
108 bp.
http://www.answers.com/topic/chromosome
Fig. 33.28 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1685232/pdf/ajhg00205-0039.pdf
11
RNA is less stable than DNA
because RNA is sensitive to
hydrolysis in basic solutions
DNA is stable in
basic solution
RNA is unstable in
base
71
12
_____________________: site-specific
Most restriction enzymes recognize
endodeoxyribonucleases causing cleavage of both
palindromes: inverted sequences with
strands of DNA at points within or near the
two-fold symmetry over two strands
specific site recognized by the enzymes; important
tools in genetic engineering
5’AAGAATTCGG3’
______________________: catalyze both
methylation of host DNA and cleavage of non- 3’ATCTTAAGCC5’
• Methylation
and restriction
at the EcoR1
site
EcoRI
GAATTC
CTTAAG
78
13
Uses of Restriction Endonucleases top
bottom
80
• DNA Fingerprinting
14