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Biotechnology Tools

SBI 4U: Molecular Genetics

What are biotech tools?

Introduction

Allow biologists to cut, join, and replicate DNA

  • Allow biologists to take specific sections of DNA and move them from one DNA molecule to another, forming recombinant DNA
  • Allow biologists to analyze sequences of DNA to determine relationships and identify organisms

Restriction enzymes

Restriction Enzymes

Endonucleases that cut DNA at specific, usually palindromic, recognition sites

  • In English: "Madam, I'm Adam"
  • In DNA: GAATTC (complementary sequence: CTTAAG)

Blunt & Sticky Ends

Blunt & Sticky Ends

Different enzymes cut DNA in different ways

  • Blunt ends: cut is made straight through DNA - no bases overhang
  • Sticky ends: cut zigzags through DNA, leaving complementary bases on one strand overhanging the other

Ligase

Two strands of DNA cut by the same restriction enzymes will have the same sticky ends

  • The same complementary bases will be exposed
  • They will form complementary base pairs with each other
  • They can be glued together, forming a new sequence... this is recombinant DNA!

What's in a Name?

Naming

Example: BamHI

  • B: Genus Bacillus
  • am: Species amyloliquefaciens
  • H: strain
  • I: Roman numeral 1 - first restriction enzyme isolated from this strain

Why don't the bacteria's enzymes cut its own DNA?

Self-Preservation

Methylases are enzymes that add a methyl group to one of the nucleotides in a recognition site, altering its chemical composition

Transgenic Organisms

Genetically Modified Organisms

  • Contain DNA from different species
  • Engineered to provide some benefit

Follow the instructions to bioengineer a (paper) bacterial plasmid that contains a gene for producing human insulin!

Paper Lab: Simulating Recombination

Glow in the Dark Animals... Why????

https://news.nationalgeographic.com/news/2009/05/shining-for-science-pictures-of-glowing-animals/#/090514-01-glowing-jellyfish_big.jpg

Glow in the Dark

  • A visible indicator of success that has no negative effects on the organism
  • Paves the way for understanding how to transfer other genes - those that might have a medical benefit to humans
  • The glow in the dark fish are sold as pets

Genetically Modified Organisms in Food

https://globalnews.ca/news/5151706/explainer-genetically-modified-food-non-gmo-labels/

Foods

Transgenic Animals

Animals

Spider Goats

Spider Goat

Silk can be used for fishing line, surgical sutures, bulletproof vests, skin repair (burn victims), etc.

Transgenic Cows

https://www.sciencelearn.org.nz/resources/834-transgenic-cows-introduction (scroll down to second video)

Polymerase Chain Reaction

A method of increasing the amount of DNA of interest (i.e. a single sequence or gene)

PCR

Applications

Molecular Identification

  • DNA Fingerprinting
  • Classification of organisms
  • Prenatal diagnosis
  • Mutation screening
  • Genetic matching
  • Detecting pathogens

Sequencing

Bioinformatics

Genomic cloning

Human genome project

Sequencing

Genetic Engineering

Site-directed mutagenesis

Gene expression studies

Genetic Engineering

Gel Electrophoresis

Method of comparing DNA from different sources

  • Produces a DNA "barcode" or "fingerprint"

Gel Electrophoresis

Gel Electrophoresis Virtual Lab

https://learn.genetics.utah.edu/content/labs/gel/

Virtual Lab

Loading the gel

Science Centre Workshop

Results

CRISPR

Origin: naturally occurring proteins in prokaryotes

  • like an immune system - detect and destroy viral DNA

Biotech application: targeted genome editing

  • Can cut out and replace specific sections of DNA

CRISPR

The Nature of Things

Make jot notes or a t-chart to list some advantages/applications and some dangers/concerns of this technology

The Genetic Revolution

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