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Introduction to molecular Introduction to molecular

biology biology


Chua Chua Kek Kek Heng Heng
Lecture/Practical Topic Lecture/Practical Topic
Introduction of Principles of Molecular Biology Introduction of Principles of Molecular Biology
Tools and Technology Tools and Technology
DNA Technology DNA Technology
Vector System Vector System
Nucleotide Sequence Nucleotide Sequence
Cloning Strategy Cloning Strategy
Gene Transfer Gene Transfer
Recombinant Clone Screening Recombinant Clone Screening
Polymerase Chain Reaction Polymerase Chain Reaction
DNA Sequencing DNA Sequencing

The purification and growth of bacteria host systems The purification and growth of bacteria host systems
Harvest of culture and plasmid preparation Harvest of culture and plasmid preparation
Quantitation Quantitation and and Agarose Agarose gel electrophoresis for plasmid DNA gel electrophoresis for plasmid DNA
PCR PCR
Ligation of DNA fragment and vector Ligation of DNA fragment and vector
Plasmid Transformation Plasmid Transformation
Recombinant clone screening using blue Recombinant clone screening using blue--white and white and Miniprep Miniprep
Restriction enzyme analysis Restriction enzyme analysis
PCR analysis PCR analysis
Assessment Assessment
Prac Prac Report (10%) Report (10%)
Conass Conass I (15%) I (15%)
Conass Conass II (15%) II (15%)
Final Exam (60%) Final Exam (60%)
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Overview Overview

DNA DNA -- Origin Origin
RNA RNA-- Expression Expression
Protein Protein-- Function Function

Structure Structure
Human genetics Human genetics
Bacterial genetics Bacterial genetics
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DNA DNA
Human Human
Contained in the nucleus Contained in the nucleus
Arranged in 22 chromosomes, plus two sex Arranged in 22 chromosomes, plus two sex
chromosomes chromosomes
Two copies of each Two copies of each


Other organisms Other organisms-- 1 or more than 1 1 or more than 1
chromosomes. chromosomes.
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5' C-G-A-T-T-G-C-A-A-C-G-A-T-G-C 3'
| | | | | | | | | | | | | | |
3' G-C-T-A-A-C-G-T-T-G-C-T-A-C-G 5'
4 Nucleotides : C G A T
Double helix
DNA Structure DNA Structure
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DNA function DNA function
Carries the blueprint for life Carries the blueprint for life
Duplication for new cells Duplication for new cells
Make proteins for biological functions: Make proteins for biological functions:
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Gene structure Gene structure
Genes: Genes:
Exons Exons, , Introns Introns
Start site Start site
Control region Control region
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mRNA mRNA
DNA is grouped into threes (codons)
AGT TTT GGG CCC AAA
Start and stop codons
mRNA is then modified......
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mRNA splicing mRNA splicing
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Alternative splicing Alternative splicing
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Translation Translation
Uses mRNA as template to make proteins Uses mRNA as template to make proteins
Occurs in Occurs in ribosomes ribosomes
One One codon codon corresponds to one amino acid corresponds to one amino acid
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Proteins Proteins
Huge proportion of cell (after H Huge proportion of cell (after H
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O) O)
Many functions: Many functions:
Structure (e.g. collagen in bone) Structure (e.g. collagen in bone)
Enzymes Enzymes
Transmembrane Transmembrane receptors receptors
Hormones Hormones
Four levels of structure Four levels of structure
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Protein structure Protein structure
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Protein interactions Protein interactions
Proteins can form interations: Proteins can form interations:
Proteins (complexes, oligomers) Proteins (complexes, oligomers)
mRNA mRNA
DNA DNA
Proteins can bind to each other depending Proteins can bind to each other depending
on their relative charges and structures on their relative charges and structures
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Common terminology Common terminology
Genome/proteome Genome/proteome
Genotype/phenotype Genotype/phenotype
Novel protein/gene Novel protein/gene
Locus/Allele/Chromosome region Locus/Allele/Chromosome region
Dominant/Recessive Dominant/Recessive
Homologous Homologous
cDNA cDNA
Dalton ( Dalton (Da Da or or kDa kDa))

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Common abbreviations Common abbreviations
SNP SNP single nucleotide polymorphism single nucleotide polymorphism
ORF ORF open reading frame open reading frame
UTR UTR untranscribed untranscribed region region
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Experimental techniques Experimental techniques
Cut/Digest, change, knockout genes Cut/Digest, change, knockout genes
Transgenic Transgenic
Sequence genes and proteins Sequence genes and proteins
Over Over--express proteins express proteins
Microarrays Microarrays
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Future of molecular biology Future of molecular biology
Personalised medicine Personalised medicine
Target Target--specific drugs (e.g. adipose tissue) specific drugs (e.g. adipose tissue)
Gene therapy Gene therapy
Comparative genomics Comparative genomics