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Molecular Biology
Dr. M. Ravichandran
Department of Medical Microbiology and Parasitology
PPSP, USM
Ext: 4592
mravic@kb.usm.my
http://www.ppsk.usm.my/lecturers/mravi/gtb204_2002_Index.htm1
History of Molecular Biology
Year Discovery Inventor
5
Prokaryotes vs Eukaryots
Prokaryotes Eukaryotes
Simple Complex
Genes grouped in Operon Genes have Introns, Exons,
Polycistronic-Multiple gene 5’cap and 3’ PolyA
No Post translational Monocistronic-Single gene
modifications Post translational
modifications
Eg: glycosylation
6
Gene expression in Prokaryots
DNA replication
(DNA polymerase)
Promoter Gene
DNA
Transcription
(RNA polymerase)
mRNA
Translation
(protein synthesis)
Protein
7
Gene expression in eukaryots
Promoter Exon Intron Exon Intron Exon
DNA
Transcription (RNA polymerase)
Primary mRNA transcript
AAAAAA
Translation
Protein
8
Electrophoresis
How do you see of DNA
DNA?
By spectrophotometer: OD 260 nm
1OD = 50ug of dsDNA
33ug of ssDNA 9
40ug of RNA
Agarose gel electrophoresis
10
Samples that have been mixed with the sample buffer is then
load into each well
11
Electrophoresis of DNA
12
Antisense molecules
Antisense molecule - RNA or DNA complementary to mRNA
Protein
mRNA
Application:
•Inhibition of HIV viral replication inside the infected cells
•Block the expression of Onchogenes Do you want to know about13RNAi
•Inhibition of tomato ripening gene in tomatoes
RNAi
14
RNA Interference (RNAi)
Double stranded RNA is
introduced into a cell
15
RISC – RNA induced silencing complex
RNA Interference (RNAi)
The pieces “guide” a
set of proteins to the
normal mRNA
16
Mechanism of RNAi
17
RNAi in zebra fish embryo
No RNA i
With RNAi
18
19
Some Facts About Biotechnology
Over 200 million people worldwide are being benefited from
biotechnology drug products and vaccines.
Biotechnology is responsible for hundreds of medical diagnostic tests
that keep the blood supply safe from the AIDS virus and detect other
conditions early enough to be successfully treated.
Biotechnology foods such as longer lasting tomatoes and better-tasting
carrots and peppers are in the market.
Hundreds of biopesticides and other agricultural products are now
used to improve our food supply and to reduce our dependence on
conventional chemical pesticides.
Environmental biotechnology products make it possible to more
efficiently clean up hazardous waste without the use of caustic chemicals or
incineration.
DNA fingerprinting, a biotech process, has dramatically improved
criminal investigation and forensic medicine, as well as afforded significant
advances in anthropology and wildlife management. 20
Suggested readings
• Principles of Gene Manipulation- An
introduction to Genetic Engineering, RW.OLD
& S.B primrose
• Genetics: TA Brown
• Molecular Biology: David Freifelder
21
Albert Einstein
22
Questions and suggestions
Dr. M. Ravichandran
Department of Medical Microbiology and
Parasitology
PPSP, USM
Ext: 4592
mravic@kb.usm.my
23