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Molecular biology application in food
biotechnology Part 1 Basic of molecular biology
Pramono, H.
outline
• Organization of the cell
• Chromosome and DNA
• Central Dogma of Molecular Biology
• Brief explanation of application of basic
molecular biology
The tree of Live
Organization of Cell
• It’s estimated that there are more than 10 million living organism in earth
• They are different, each reproduces it self faithfully progeny of same species
• This phenomenon is called heredity stored information in linear DNA form
• Most living organism are single or multicellular– Prokaryotic cell
– Eukaryotic cell
Inside the cell
Cell membrane
• Consist of protein membrane,
transmembrane, and phospholipid bilayer
• Keep homeostatis of the cell
Cell wall
Organelles
Organelles Function Ribosome Most abundant organelle, translation of
mRNA
Mitochondria Energy generating organelle in Eukaryotic
cell
Plasmid Small circular DNA fragment in
bacteria or yeast, some specific
function
Chromosome Linear DNA for generating genetic
information heredity
Reticulum endoplasmic Post translational modification of protein
Golgi complex Post translational modification and
generate liposome to bring molecule
exclude from the cell
Chromosome
DNA
• Information always stored as code– Language: consist of alphabetic code and voice
– Music in CD or Flash: data consist of 1 and 0 code
– Live: DNA fragment called gene
• DNA consist of phosphate, sugar, and
nitrogen base nucleotide
• Double helix (Watson and Crick)
• Note: 5 (phosphate)-3 (OH)
Denaturation of DNA
• Hidrogen bond between nitrogen base of
DNA can denaturated when high temperature
occurs
• GC content impact Tm (melting point) of DNA
• Denaturation of DNA or RNA can be deduced
by λ260
– Double helix DNA A260 :1.0
– Single strand DNA A260 :1.37
– Free Nucleotide A260 :1.60
Tm
• Tm important for application in PCR
• Tm affected by
– G-C content (3 hidrogen bond)
– Ionic of solution (remember that DNA is
negative charged) NaCl increase, Tm
increase
Central Dogma
Replication
• Replication is forming new DNA strand
from old or parental DNA that exist inside
the cell
• Mechanism:
– form DNA replication fork
– Start from ORI (origin of replication)
– 5-3 direction
Replication fork model
Transcription
• Occurs by action of RNA polymerase in chromosome fragment called promoter
• Splicing occurs in Eukaryotic mRNA processing
• Cell produces several type of RNA
– miRNA microRNA
– siRNA small interfering RNA
– tRNA transfer RNA
– rRNA ribosomal RNA
– mRNA messenger RNA
Promoter and Operon in
Prokaryotic cell
quizlet.com
Transcription in Eukaryotic cell
www.phschool.com
Translation
• Translate gene into protein for many
purposes
– Generating enzyme
– Cellular signaling (ex: hormone)
– Regulating metabolism and cell
• Translate nucleotide protein
Translation code
Brief Application
• Organization of the cell:– Cell wall disruption by chloroform and enzyme
• Chloroform disrupt cell membrane
• Proteinase K degrade protein transmembrane
• Lisozyme degrade cell wall (peptidoglycan)
• Chromosome and DNA:– Isolation of chromosomal DNA, gene
– Molecular detection
– Transformation of DNA to other bacteria
• Central Dogma of Molecular Biology:– Cloning and PCR
– Expression of gene in model organism
Any question?
