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

Splicing intron

chemistry.umeche.maine.edu

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?