Biotechnology

Biology- Regular

John Murnan

Etowah High School

Welcome to Biotech Inc.

Your new job requires some tools

• Biotech Inc. does research into developing products for Genetic Engineering, DNA fingerprinting and Cloning

• As interns you will learn how to use some new tools and techniques

• Please open your tool box and lets see what’s inside it.

Restriction Enzymes

– Act like scissors– Cut DNA at specific places

Recognition sites of different restriction enzymes

Restriction site function

Function of

restriction enzymes

Ligase

• Acts like glue

• Connects two pieces of DNA– Makes bond between sugar-phosphate on

backbone.

DNA Polymerase

• Acts like a copier– Copies a strand of DNA

• Requires free nucleotides to make the copies

Retroviruses

• Can insert DNA into a chromosome in a living cell– HIV, the virus that

causes AIDS is one type.

Some smaller tools

• Very thin syringe– Used to remove nucleus from cells

• Micropipette– A very small eyedropper– Used to move very small amounts of fluid

Gel Electrophoresis

• Sorts molecules by size– Uses a gel like jello and an electric current.

Gel Electrophoresis Setup

Gel Electrophoresis

Pictures of gel electrophoresis

PCR

• Uses DNA Polymerase and free nucleotides to make millions of copies of DNA from a single strand.

Overview of PCR

Effect of PCR

Genetic Engineering

Manipulating organisms to create products

• Use restriction enzymes, ligase, Gel Electrophoresis and PCR

• Inserts foreign genes into bacteria– Makes bacteria produce proteins– Bacteria can be used to store genes

Plasmid Ligation

• Puts DNA from humans (or other species) into bacteria DNA.

Review of Bacteria Structure

• Nucleoid Region- is bacteria chromosome

• PLASMID- small rings of DNA that have only a few genes.

Plasmid

Ligation of plasmid 1

Ligation of plasmid 2

Genetic Engineering with plasmid

Possible products of

genetic engineering

Using Plasmid Ligation techniques, we can make both proteins and genes that can be used to

DNA Fingerprinting

Forensic Scientists

• Use restriction enzymes, Gel Electrophoresis, PCR

• Can identify any person based on their DNA sequences.

• Each of us has unique sequences.

What is an RFLP?

Sometimes mutations occur which cause recognition sites to be changed.

If a person has a mutation, then the site is changed and DNA of different lengths is produced using the same restriction enzyme.

Restriction Fragment Length Polymorphism

Why you are different than other people

Southern Blot 1

Southern blot results

DNA fingerprinting

example

• http://www.koshlandsciencemuseum.org/exhibitdna/index.jsp

Cloning

Cloning Activity

• http://gslc.genetics.utah.edu/units/cloning/clickandclone/

Ethics of cloning

• Occurs naturallly (identical twins)

• Easier screening of embryos for genetic diseases

• Increase chances for infertile couples to have children

• High risk of developmental errors leading to miscarriages

• Cells are as old as the individual they are taken from

Gene Therapy

Gene therapy example

Gene therapy vectors

Delivery methods

Gene Therapy example