Observe the following slide and: 1) Explain what is occurring on the right side and then on the left side. 2) What might be the purpose of doing the process on the left?

Human Gene Plasmid DNA Human protein Completed file! + +

Producing Insulin in bacteria Human protein in Cow milk! Pest-resistant Corn

Purposes, for example: 1. Make bacterial factories of medicines. 2. Produce genetically improved plants and animals 3. Insert correcting genes into patients for Gene therapy. Notes

Recombinant DNA Technology Or Genetic Engineering

Student Guide Lab 2a

How do we insert a foreign DNA into an organism? 1. Extraction and restriction 2. Building a construct of mixed DNA origins (Recombination) 3. Delivering into the organism (Transformation) Notes

It is as simple as a Lego game!

1. Extraction and cleavage (restriction) 2. Combining gene of interest with vector DNA (e.g. plasmid) = “recombination”. 3. Delivering into the organism (Transformation)

Recognition sequences Restriction enzyme EcoRI cuts the DNA into fragments. Sticky end

Originally protected bacteria from viruses. Restriction Site = Recognition sequence NOT Cleavage site Recognition site is a planidrome (just like “radar” “Madam, I’m adam”

Palindromes:

Genetic Engineering & Working with DNA DNA Restriction - Animation from DNA Learning Center

To the students presenting: The following slides are NOT part of your presentation, but for context only.

1. Extraction and cleavage (restriction) 2. Combining gene of interest with vector DNA (e.g. plasmid) = “recombination”. 3. Delivering into the organism (Transformation)

Plasmid DNA PLASMID = A circular DNA that easily gets into bacterial cells, and replicates independently. Notes

Bacteria naturally pass plasmids to one another by conjugation. Result: Transformation of the accepting cell. Conjugation animation Notes

Plasmid of our interest: pARA-R Gene of interest Selection markers Origin of replication

How was this plasmid constructed?

Sticky ends Recombinant DNA Human Cell Gene for human insulin Bacterial cell with gene for human growth hormone Bacterial Chromosome Plasmid

1. Extraction and cleavage (restriction) 2. Combining gene of interest with vector DNA (e.g. plasmid) = “recombination”. 3. Delivering into the organism (Transformation)

Transformation = Inserting foreign DNA into a living cell. Bacterial reproduction Animation

Exercise: Constructing paper ‘plasmid’ that contains the gene for human ‘insulin’. Day1: * Cut out the DNA strips, connect them: Gene – in order. Plasmid – in any particular order. * Mark all the 9 restriction and cleavage sites.