Recombinant DNA TechnologiesRecombinant DNA Technologies

A.DNA- Deoxyribonucleic AcidA.DNA- Deoxyribonucleic Acid1.1. Bases:Bases:

a.a. A- AdenineA- Adenine

b.b. C- CytosineC- Cytosine

c.c. G- GuanineG- Guanine

d.d. T- ThymineT- Thymine

-put together in a double-helical molecule-put together in a double-helical molecule

with A-T & C-G as the “rungs”with A-T & C-G as the “rungs”

-form GENES-form GENES

e. We have about 30,000 genes and they are e. We have about 30,000 genes and they are mapped by location on each chromosomemapped by location on each chromosome

-”Human Genome Project”-”Human Genome Project”

f. We are 99.9% identical; .1% makes us unique f. We are 99.9% identical; .1% makes us unique and different from Rob Marder (thank heaven!)and different from Rob Marder (thank heaven!)

B. DNA fingerprinting B. DNA fingerprinting

1. Find areas of variation1. Find areas of variation

2. Hit with restriction enzymes2. Hit with restriction enzymes=>neat DNA cutting tools (listen for where =>neat DNA cutting tools (listen for where

they came from!)they came from!)

3. Run through a gel 3. Run through a gel (electrophoresis)(electrophoresis)

a. After cutting, small particles move a. After cutting, small particles move fast, larger ones move slowlyfast, larger ones move slowly

b. Gives a banding pattern:b. Gives a banding pattern:

Gel Electrophoresis & Apparatus

Two examples to try:

-Crime Scene & Paternity

Testing (Maury Povich style!)

C. Gene TherapyC. Gene Therapy1. Problems with gene expression lead to faulty 1. Problems with gene expression lead to faulty molecules, especially enzymesmolecules, especially enzymes

-cannot do their jobs-cannot do their jobs

2. examples: Cystic Fibrosis & Tay Sachs 2. examples: Cystic Fibrosis & Tay Sachs diseasesdiseases

3. Idea: replace bad genes with good ones that 3. Idea: replace bad genes with good ones that make the make the proper moleculeproper molecule

And theoretically “fix” the problemAnd theoretically “fix” the problem

4. How can we deliver the new genes?4. How can we deliver the new genes?

(i.e. what do we know that inserts genes over existing (i.e. what do we know that inserts genes over existing ones…. Think “poison”)ones…. Think “poison”)

Called “vectors”Called “vectors”

D. CloningD. Cloning1.1. Genetically identical copiesGenetically identical copies

2.2. Replace nucleus of egg w/ compat. Adult Replace nucleus of egg w/ compat. Adult DIPLOID nucleus and jolt w/ electricityDIPLOID nucleus and jolt w/ electricity

3.3. Age of clone- new or as old as donated Age of clone- new or as old as donated DNA?DNA?

4.4. Just because we can, does that mean we Just because we can, does that mean we should?should?

5.5. Examples:Examples:

E. PCR- Polymerase Chain ReactionE. PCR- Polymerase Chain Reaction

1.Takes small 1.Takes small amounts of DNA amounts of DNA and copies it and copies it millions of timesmillions of times

2. Useful when DNA 2. Useful when DNA quantities are quantities are minuteminute

-crime scenes-crime scenes

-archeological-archeological

findsfinds

F. Transgenic organismsF. Transgenic organisms

1. Organisms with foreign DNA spliced into it

2. Examples:

a. Bacteria- make chemicals that we need

1) insulin (rather than from a dead pig’s organ)

2) fertilizers

3) hormones

4) nutrasweet (phenylalanine- watch out PKU people)

5) Spider silk for manufacturing

b. Plants

1) Resist frost (Arctic flounder gene)

2) Disease resistance

Transgenic organisms continued:Transgenic organisms continued:

c. Animalsc. Animals

1. Goats that produce anti-1. Goats that produce anti-clotting proteinclotting protein

2. 2. GlowingGlowing Rabbits: Rabbits:

-Fun -Fun , but actually are, but actually are

““markers”markers”

3. “Human” Mice3. “Human” Mice

Benefit?Benefit?

4. Rejection –resistant pigs 4. Rejection –resistant pigs for organ transplants into for organ transplants into

humans humans ( (xenotransplantationxenotransplantation))

Example of how we generate HUMAN insulin from bacteria:

•What are some benefits to utilizing this method?

•How did Diabetics get their insulin before this was developed?

•Hint: Our friend, the pig

Genetic engineering funniesGenetic engineering funnies