Genetic engineering
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Genetic Engineering. Intent of altering human genome Introducing new genetic material into genome Isolating genes to produce on large scale ( Insulin). Recombinant DNA. DNA that contains genes of two species How? Restriction enzymes – cut out desired gene

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Genetic Engineering

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Genetic engineering

Genetic Engineering

  • Intent of altering human genome

  • Introducing new genetic material into genome

  • Isolating genes to produce on large scale (Insulin)


Recombinant dna

Recombinant DNA

  • DNA that contains genes of two species

  • How?

    • Restriction enzymes – cut out desired gene

    • Occur naturally in prokaryotic cells

    • Recognize specific recognition sites – 4 to 8 base pairs

    • Recognition sites are palindromes

    • Cuts gene (digests) in one direction only

    • Creates restriction fragments


Process restriction enzyme

Process – Restriction Enzyme

  • Locates recognition site (Top Strand)

  • Cuts the DNA backbone

  • Locates recognition site (Bottom Strand)

  • Cuts the DNA backbone

  • DNA separates


Sticky ends or blunt ends

Sticky Ends or Blunt Ends

  • Sticky ends – zigzag cuts in strand

  • Blunt ends – straight cut across strand


Putting fragments together

Putting Fragments together

  • DNA ligase – sticky ends

  • T4 DNA ligase – blunt ends

    • forms phosphodiester bonds in DNA


Plasmids

Plasmids

  • Small circular pieces of DNA found in bacteria

  • Used as vectors for recombinant DNA (artificial)

  • Restriction enzymes used to isolate specific gene are used to cut plasmids


Process

Process

  • Plasmids and DNA fragments are placed in same solution

  • Anneal

  • DNA ligase is used to form phosphdiester bond

  • Recombinant DNA introduced into host cell

  • DNA is cloned


Identifying bacteria clones with target genes

Identifying Bacteria Clones with Target Genes

  • Hybridization – identify cells that contain recombinant DNA

  • Identified using a hybridization probe – short single stranded complementary DNA molecule

  • Once identified bacteria can be grown in huge quantities (commercial use)


Transformation

Transformation

  • Cells that receive foreign DNA

  • Bacterial cells sometimes will not take up a plasmid

  • Bacteria are placed in ice water bath containing CaCl₂

  • Solution is heated and cooled repeatedly disrupting plasma membrane of bacteria allowing plasmid to enter

  • Solution is kept at 37⁰C to stabilize and grow


Cloning insulin

Cloning Insulin


Restriction maps

Restriction Maps

  • Diagrams that show all recognition sites on a specific plasmid and distances in base pairs

  • Shows which restriction enzyme should be used

  • Allow scientists to determine which plasmids will work the best for cloning experiments


Gel electrophoresis

Gel Electrophoresis

  • Technique used to separate fragments of DNA (PCR)

  • Used to help identify information about unknown pieces of DNA (restriction maps/forensics)


Genetic engineering

PCR

  • Polymerase Chain Reaction

    • Increase number of DNA copies from a single biological sample in a few hours

    • Only specific regions of a chromosome are replicated

  • Process

    • Denaturation

    • Annealing

    • Elongation


Genetic engineering

  • Taq polymerase is used to put strand together

  • Isolated from bacteria that live in hot springs


Genetic engineering1

Genetic Engineering

  • Biopharming

    • Pharmaceutical products produced on large scale

    • Organisms are genetically engineered to produce a specific protein

    • Ability to make new protein is passed on to offspring


Genetic engineering2

Genetic Engineering

  • Transgenic Organism (genetically modified organism, GMO)

    • Organisms that contain one or more genes from another organism


Knockout mice

Knockout Mice


Why use genetic engineering

Why use Genetic Engineering?

  • Cost

  • Larger organisms can produce larger molecules

  • Better versions of organisms

  • 80% of Canadas Canola crop is GM


Gene therapy

Gene Therapy

  • Techniques used to replace, remove or alter a defective gene before symptoms are expressed

  • Insert healthy genes into cells of tissues that are affected by a disorder


Gene therapy1

Gene Therapy


Gene therapy2

Gene Therapy

  • Germ-line gene therapy

    • Genes introduced in sperm or egg cells

    • Passed on to future generations

  • Somatic gene therapy

    • Genes introduced into body cells (not sperm or egg)

    • Will not be passed on


Biotechnology

Biotechnology

  • Artificial Insemination

    • Sperm is collected and concentrated before being placed in woman’s uterus

  • In Vitro Fertilization

    • Eggs from mom are retrieved and combined with sperm in petri dish

    • After fertilization, zygote is placed in woman’s uterus


Genetic testing

Genetic Testing

  • Determining if your baby carries a gene that is associated with an increased risk of developing a disease


Aminocentesis

Aminocentesis

  • Benefits

    • Diagnosis of chromosome abnormalities

    • Diagnosis of other defects

  • Risks

    • Discomfort for expectant mother

    • Slight risk of infection

    • Miscarriage


Chorionic villus sampling

Chorionic Villus Sampling

  • Benefits

    • Diagnosis of chromosome abnormalities

    • Diagnosis of certain genetic defects

  • Risks

    • Miscarriage

    • Infection

    • Newborn having limb defects


Genetic engineering

Stem Cells


Cord blood banking

Cord Blood Banking

  • Source of hematopoietic (blood forming) stem cells

  • Blood (stem cells) found in umbilical cord after birth

  • Similar to tissue stem cells

  • Develop into various blood cells

  • These stem cells could be banked (stored)

  • Used to treat variety of diseases


Genetic engineering

http://www.youtube.com/watch?v=8JTw2RpDo9o&feature=related

http://www.youtube.com/watch?v=VOXJaIvRltM

http://www.youtube.com/watch?v=czzCmgr0IgQ&feature=related

http://www.youtube.com/watch?v=VOXJaIvRltM

http://www.youtube.com/watch?v=qiQfGBOoMPM&feature=fvst

http://www.youtube.com/watch?v=w9wIxe4WGFg&feature=related

http://www.youtube.com/watch?v=L-Rj_rvBLZ0&feature=related

http://www.youtube.com/watch?v=-FaGf0h20Vw


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