Chapter 20: Terms to Know

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Chapter 20 Terms to Know

1. Genetic engineering
2. Biotechnology
3. Recombinant DNA
4. Gene cloning
5. Restriction enzymes
6. Sticky ends
7. DNA ligase
8. Cloning vector
9. Nucleic acid hybridization
10. Genomic library

• cDNA library
• PCR
• Gel electrophoresis
• Southern blotting
• DNA microarray assays
• SNPs
• RFLPs
• Stem cells
• Gene therapy
• GMO (genetically modified organism)

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What You Must Know

• The terminology of biotechnology.
• The steps in gene cloning with special attention
  to the biotechnology tools that make cloning
  possible.
• The key ideas that make PCR possible.
• How gel electrophoresis can be used to separate
  DNA fragments or protein molecules.

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• Genetic Engineering process of manipulating
  genes and genomes
• Biotechnology process of manipulating organisms
  or their components for the purpose of making
  useful products.

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• Recombinant DNA DNA that has been artificially
  made, using DNA from different sources
• eg. Human gene inserted into E.coli
• Gene cloning process by which scientists can
  product multiple copies of specific segments of
  DNA that they can then work with in the lab

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Tools of Genetic Engineering

• Restriction enzymes (restriction endonucleases)
  used to cut strands of DNA at specific locations
  (restriction sites)
• Restriction Fragments have at least 1 sticky end
  (single-stranded end)
• DNA ligase joins DNA fragments
• Cloning vector carries the DNA sequence to be
  cloned (eg. bacterial plasmid)

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Using a restriction enzyme (RE) and DNA ligase to
make recombinant DNA
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Gene Cloning
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Applications of Gene Cloning
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Techniques of Genetic Engineering
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Techniques of Genetic Engineering

• Transformation bacteria takes up plasmid (w/gene
  of interest)
• Nucleic acid hybridization used to track gene of
  interest
• PCR (Polymerase Chain Reaction) amplify (copy)
  piece of DNA without use of cells
• Gel electrophoresis used to separate DNA
  molecules on basis of size and charge using an
  electrical current (DNA ? pole)
• Southern blotting used to find a specific human
  gene
• DNA microarray assays study many genes at same
  time

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PCR (Polymerase Chain Reaction) amplify (copy)
piece of DNA without use of cells
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Nucleic Acid Hybridization find and track gene
of interest
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Gel Electrophoresis used to separate DNA
molecules on basis of size and charge using an
electrical current (DNA ? pole)
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Gel Electrophoresis used to separate DNA
molecules on basis of size and charge using an
electrical current (DNA ? pole)
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Southern Blotting used to find a specific gene
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Microarray Assay used to study gene expression
of many different genes
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DNA microarray that reveals expression levels of
2,400 human genes
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Cloning Organisms

• Nuclear transplantation nucleus of egg is
  removed and replaced with nucleus of body cell

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Nuclear Transplantation
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Problems with Reproductive Cloning

• Cloned embryos exhibited various defects
• DNA of fully differentiated cell have epigenetic
  changes

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Stem Cells

• Stem cells can reproduce itself indefinitely and
  produce other specialized cells
• Zygote totipotent (any type of cell)
• Embryonic stem cells pluripotent (many cell
  types)
• Adult stem cells multipotent (a few cell types)
  or induced pluripotent, iPS (forced to be
  pluripotent)

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Embryonic vs. Adult stem cells
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Using stem cells for disease treatment
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Applications of DNA Technology

1. Diagnosis of disease identify alleles, viral
   DNA
2. Gene therapy alter afflicted genes
3. Production of pharmaceuticals
4. Forensic applications DNA profiling
5. Environmental cleanup use microorganisms
6. Agricultural applications - GMOs

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Gene therapy using a retroviral vector
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Pharm animal produce human protein secreted in
milk for medical use
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DNA Fingerprinting
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RFLPs (rif-lips)

• Restriction Fragment Length Polymorphism
• Cut DNA with different restriction enzymes
• Each person has different s of DNA fragments
  created
• Analyze DNA samples on a gel for disease
  diagnosis
• Outdated method of DNA profiling (required a
  quarter-sized sample of blood)

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RFLPs Disease Diagnosis
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STR Analysis

• STR Short Tandem Repeats
• Non-coding DNA has regions with sequences (2-5
  base length) that are repeated
• Each person has different of repeats at
  different locations (loci)
• Current method of DNA fingerprinting used only
  need 20 cells for analysis

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STR Analysis
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STR Analysis
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Biotechnology Techniques
How to make Recombinant DNA Gel Electrophoresis PCR
Summarize What is this technique?
Draw and label a diagram to show this technique
What are the main tools or materials involved?
Applications What is this being used for?