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Basic concepts of
Genes, DNA & RNA
Course: BSc Biotech Sem-I
Subject : Fundamentals of Biotechnology
Unit - 2
1
BREAKTHROUGH DISCOVERY
• In 1953, James Watson and Francis Crick
discovered the double helical structure of the
DNA molecule
2
The discovery was announced on
February 28, 1953
3
The First Paper appeared in Nature on April
25, 1953
4
5
Watson and Crick - Awarded Nobel Prize
in 1962
6
DNA
A purine always links with a pyrimidine base to maintain the structure of DNA.
Adenine ( A ) binds to Thymine ( T ), with two hydrogen bonds between them.
Guanine ( G ) binds to Cytosine ( C ), with three hydrogen bonds between them.
7
8
Chemical Structure Of DNA
9
10
Nucleoside & Nucleotide, the
fundamental building block of DNA
glycosidic bond
phosphoester bond
11
12
13
Ribose
14
DNA & RNA - Nucleotide Bases
15
Nucleotide
adenine
deoxyribose
PO4
16
Nucleotides
17
14.
Chargaff's rule
• Chargaff's rules state that DNA from
any cell of all organisms should have a 1:1
ratio (base Pair Rule)
of pyrimidine and purine bases and, more
specifically, that the amount of guanine is
equal to cytosine and the amount
of adenine is equal to thymine.
• They were discovered by Austrian
chemist Erwin Chargaff
18
Relative proportions (%) of bases in DNA
Organism %A %G %C %T A/T G/C %GC %AT
φX174 24 23.3 21.5 31.2 0.77 1.08 44.8 55.2
Maize 26.8 22.8 23.2 27.2 0.99 0.98 46.1 54
Octopus 33.2 17.6 17.6 31.6 1.05 1 35.2 64.8
Chicken 28 22 21.6 28.4 0.99 1.02 43.7 56.4
Rat 28.6 21.4 20.5 28.4 1.01 1 42.9 57
Human 29.3 20.7 20 30 0.98 1.04 40.7 59.3
Grasshoppe
r 29.3 20.5 20.7 29.3 1 0.99 41.2 58.6
Sea Urchin 32.8 17.7 17.3 32.1 1.02 1.02 35 64.9
Wheat 27.3 22.7 22.8 27.1 1.01 1 45.5 54.4
Yeast 31.3 18.7 17.1 32.9 0.95 1.09 35.8 64.4
E. coli 24.7 26 25.7 23.6 1.05 1.01 51.7 48.3
19
CHEMICAL AND PHYSICAL PROPERTIES
OF DNA
• ABSORPTION
• The bases in DNA absorb ultraviolet light at
the wavelength of 260 nm
• This absorption can be monitored using a
spectrophotometer
• This is one method used to figure the
concentration of DNA in solution
• The more DNA present, the higher the
absorption
20
• DENSITY
• Density can be measured by CsCl-density
ultracentrifugation
• can be used to estimate G+C content
• GC base pairs are more dense than AT base
pairs
• Density studies show the existence of satellite
DNA
21
• DENATURATION
• DNA is considered denatured when the double
stranded DNA molecule is converted into two
single stranded molecules
• As thermal energy increases, the frequency of
hydrogen bonds breaking between the molecules
increases
• G-C base pairs are held together by three
hydrogen bonds (A-Ts by two) and it therefore
takes more energy (higher temperatures) to
separate molecules with high GC contents
22
• Hydrophobicity of solvent
• Hydrophobic substances will allow the bases in
DNA to dissolve into the solvent
• Whereas hydrophilic substances will keep the
bases of DNA stacked upon one another in
the orientation that most favors hydrogen
bonding between DNA strands
23
• pH
• Acidic pH cause breakage of phosphodiester
bonds between nucleotides and breakage of
the N-glycosidic bond between the sugar and
purine bases
• Alkali - Above pH 11.3, all hydrogen bonds
are disrupted and the DNA is totally
denatured
• Salts will stabilize the DNA double helix
24
• Electrophoresis
• DNA has a negative charge that is proportional to
its size
• This is due to the negatively charged phosphates
in the sugar-phosphate backbone
• If DNA is placed in an electrical field it will migrate
towards the positive electrode (the cathode)
• smaller pieces will migrate faster than larger pieces
• Larger pieces have trouble squeezing through the
gel matrix and are hence retarded while smaller
pieces migrate easier
25
• Type of gels
• Agarose is used to separate fairly large DNA
molecules
– 5 million to a few thousands base pairs
• Polyacrylamide is used to separate small pieces
of DNA
– 2 to several hundred base pairs
• The size of DNA is estimated by comparing
its migration through the gel to DNA
molecules of known size
26
A, B and Z DNA
• A form – favored by
RNA
• B form – Standard
DNA double helix under
physiological conditions
• Z form – laboratory
anomaly,
– Left Handed
– Requires Alt. GC
– High Salt/ Charge
neutralization
27
15.
28
16.
29
17
RNA
Three major classes of RNA:
Difference between RNA & DNA
RNA DNA
RNA nucleotides contain ribose sugar DNA contains deoxyribose
RNA has the base uracil DNA has the base thymine
presence of a hydroxyl group at the 2'
position of the ribose sugar.
Lacks of a hydroxyl group at the 2'
position of the ribose sugar.
RNA is usually single-stranded DNA is usually double-stranded 30
mRNA
• Transcripts of structural genes.
• Encode all the information necessary for the
synthesis of a polypeptide of protein.
• Intermediate carrier of genetic information;
deliver genetic information to the cytoplasm.
3118
mRNA to Amino Acid Dictionary
32
19
33
20
tRNA
 RNA molecules 70- 100 nucleotide long.
 The secondary structure of the tRNA
resembles a D loop, anticodon loop,
and T loop and the acceptor stem.
 Carry correct amino acids to their
position along the mRNA template to be
added to the growing polypeptide chain.
34
21
rRNA
• The central component of the
ribosome.
• Ribosome; factory for protein
synthesis; composed of ribosomal
RNA and ribosomal proteins.
• rRNA provides a mechanism for
decoding mRNA into amino acids.
• rRNA interact with the tRNAs during translation by
providing peptidyl transferase activity.
35
22
RNA Helices are short, bulges, loops
36
23
Organization of DNA in eukaryotes
37
24
• Chromosome –
– double stranded DNA
molecule packaged by histone
& scaffold proteins
DNA double helix
nucleosome
30nm fiber
condensed chromosome
38
25
Genome
Gene
• Is the basic units of
inheritance; it is a segment
within a very long strand of
DNA with specific
instruction for the
production of one specific
protein.
• Genes located on
chromosome on it's place or
locus.
Genome and Gene
• Totality of genetic information of an organism.
• Encoded in the DNA (for some viruses, RNA).
39
26
Image refrences
1.http://www.bbc.co.uk/history/historic_figures/crick_and
_watson.shtml
2. http://m.harunyahya.com/tr/works/4159/Once-Upon-A-
Time-There-Was-Darwinism/chapter/4732/Once-there-
was-no-knowledge-of-biological-information
3. http://arabian-explorers.com/james-watson-francis-crick
4.https://www.bcps.org/offices/science/secondary/forensi
c/DNA.pdf
5.http://www.timerime.com/en/event/209438/Nobel+Prize
+for+Literature/
6. http://pixgood.com/dna-model-labeled-structure.html
40
Image refrences
7.http://www.yourarticlelibrary.com/dna/deoxyribonucleic
-acid-dna-model-chemical-composition-and-
transformation-experiments/22676/
8.http://ircamera.as.arizona.edu/NatSci102/NatSci/text/nuc
leicacid.html
9. http://quizlet.com/11685525/chapter-8-slides-flash-cards/
10.http://biology200.gsu.edu/houghton%2704/4564%20%27
14/lecture6.html
11. http://www.slideshare.net/oreonixha/dna-11921134
12. http://www.bvgh.org/Current-Programs/Neglected-
Disease-Product-Pipelines/Global-Health-
Primer/Targets/cid/ViewDetails/ItemID/18.aspx
41
Image refrences
13. http://smarttechno.net/tag/purine-bases
14. http://en.wikipedia.org/wiki/Nucleotide
15.http://biomedical.youngdong.ac.kr/sgkim/files/2013/06/P
OB_5e_ch08-SGKim.pdf
16. https://lh3.ggpht.com/b7SCiLAza-
c_D_wbjzOP86F4AnXyKu_wrviHfbF3GFqCWJXzuqHQvIG
7P51jXvJcNJnNfQ=s144
17.https://lh5.ggpht.com/uE2KYozDJ302Z1ScuRJwtmqjjthxv
ILPxYnYyxog8q75VSahpvcMgtztJl8qGrXKhgsugA=s136
18. https://lh4.ggpht.com/1cWohlnifUAe4-
WcqCCzkt3_GjfAmlbW1Tc7D0rcQXm9j5qW6Ns_xRv11VP
wLy1aOsRkaQ=s170
42
Image refrences
19.https://lh4.ggpht.com/u0_RqejooRi2Vz7xOVJ7UkYuzUZquHOQ
nm4hSncWyNEOF0nAvJcM4WQlkYPuWaf2JiFN=s101
20. https://lh3.ggpht.com/ma2QHNXrMmoFjD-
RtySlsVEPZjKqVEh7dv2oytsVIXO570HJZUXABdFoleQeALL8wy1f
bg=s85
21.https://lh4.ggpht.com/rp027AgQNGhK99zk9t5qoLYRJgES8MDj
0tnom_OYqQbMbTepfE1UttFktW5mSDBKuzUQ=s85
22.https://lh4.ggpht.com/iFsbqDCWZz_IqdhnffJBMcOFPvy60Nzd
BMG9V8SF4hIzzPYpgOJj4uhhnOct-vCGCB1k=s85
23. http://people.csail.mit.edu/donglai/
24. http://www.jeansforgenesday.org/disorders/basicterms
25. http://carolguze.com/text/442-1-humangenome.shtml
26. http://science.howstuffworks.com/life/cellular-
microscopic/cell7.htm
43
References
• Biochemistry by S C Rastogi
• Genes XI by Krebs, Goldstein and Kilpatrick
• Biotechnology by H K Das
• Biotechnology by John E. Smith (5th Ed.)
• http://www.genomicglossaries.com/
• http://biology-pages.info
44

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B sc biotech i fob unit 2 gene, dna and rna

Hinweis der Redaktion

  1. Nucleosides consist of a base (pyrimidine or purine) attached to a sugar (ribose or deoxyribose). Nucleotides are nucleoside phosphates Nucleotides may be nucleoside monophosphates, diphosphates or triphosphates or NMPs, NDPs, and NTPs, respectively
  2. The vast majority of the DNA molecules present in the aqueous protoplasms of living cells almost certainly exist in the Watson-Crick double helix form described above. This is called the B-form of DNA and shows right-handed coiling. It contains 10.4 base pairs per turn (instead of the 10 men­tioned above). Dehydrated DNA occurs in the A-form which is also a right handed helix, but it has 11 base pairs per turn. Certain DNA sequences occur in Z-form, which shows left-handed coiling, contains 12 base pairs per turn. In Z-DNA, the sugar-phosphate backbone follows a zig-zagged path giving it the name Z-DNA or Z-form. Specific segments of a DNA molecule can undergo conformational changes from B-form to Z-form and vice-versa; these changes may be brought about by some specific regulatory proteins. The Z-form DNA is postulated to play a role in gene regulation.
  3. A nucleosome is composed of DNA & histones A nucleosome wraps approx. 200bp DNA