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Chapter 20: Terms to Know

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Chapter 20: Terms to Know Genetic engineering Biotechnology Recombinant DNA Gene cloning Restriction enzymes Sticky ends DNA ligase Cloning vector Nucleic acid ... – PowerPoint PPT presentation

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Title: Chapter 20: Terms to Know


1
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)

2
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.

3
  • Genetic Engineering process of manipulating
    genes and genomes
  • Biotechnology process of manipulating organisms
    or their components for the purpose of making
    useful products.

4
  • 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

5
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)

6
Using a restriction enzyme (RE) and DNA ligase to
make recombinant DNA
7
Gene Cloning
8
Applications of Gene Cloning
9
Techniques of Genetic Engineering
10
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

11
PCR (Polymerase Chain Reaction) amplify (copy)
piece of DNA without use of cells
12
Nucleic Acid Hybridization find and track gene
of interest
13
Gel Electrophoresis used to separate DNA
molecules on basis of size and charge using an
electrical current (DNA ? pole)
14
Gel Electrophoresis used to separate DNA
molecules on basis of size and charge using an
electrical current (DNA ? pole)
15
Southern Blotting used to find a specific gene
16
Microarray Assay used to study gene expression
of many different genes
17
DNA microarray that reveals expression levels of
2,400 human genes
18
Cloning Organisms
  • Nuclear transplantation nucleus of egg is
    removed and replaced with nucleus of body cell

19
Nuclear Transplantation
20
Problems with Reproductive Cloning
  • Cloned embryos exhibited various defects
  • DNA of fully differentiated cell have epigenetic
    changes

21
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)

22
Embryonic vs. Adult stem cells
23
Using stem cells for disease treatment
24
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

25
Gene therapy using a retroviral vector
26
Pharm animal produce human protein secreted in
milk for medical use
27
DNA Fingerprinting
28
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)

29
RFLPs Disease Diagnosis
30
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

31
STR Analysis
32
STR Analysis
33
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34
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?
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