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Introduction to biotechnology

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Introduction to biotechnology Haixu Tang School of Informatics Biotechnology Cell technology Isolating cells Growing cells in culture Fractionating cells Molecular ... – PowerPoint PPT presentation

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Title: Introduction to biotechnology


1
Introduction to biotechnology
  • Haixu Tang
  • School of Informatics

2
Biotechnology
  • Cell technology
  • Isolating cells
  • Growing cells in culture
  • Fractionating cells
  • Molecular technology
  • DNA cloning
  • DNA sequencing
  • Gene expression
  • Analyzing protein functions

3
Isolating Cells
  • Disrupting extracellular matrix proteolytic
    enzymes or EDTA
  • Separating different cell types
  • Antibodies coupled to a fluorescent dye
    (fluorescence-activated cell sorter)
  • Microscopic dissection
  • Cell growth

4
Fluorescence-activated cell sorter
5
Microscopic dissection
6
Growing cells in culture
7
Composition of a Typical Medium Suitable for the
Cultivation of Mammalian Cells
8
Cell line
  • Most vertebrate cells stop dividing after a
    finite number of cell divisions in culture
    senescence
  • "immortalized" cell line telemerase
  • Inactivate the checkpoint mechanisms
  • Cell lines can often be most easily generated
    from cancer cells.

9
Hybrid cells
  • a heterocaryon, a combined cell with two separate
    nuclei

10
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11
Ultracentrifuge
12
Cell fractionation by centrifugation
13
velocity sedimentation vs. equilibrium
sedimentation
14
Column chromatography
15
Matrices used for chromatography
16
Protein purification by chromatography
17
SDS polyacrylamide-gel electrophoresis
18
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19
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20
Western blotting
21
Protein identification via MS
22
DNA recombination technology
  • 1. Cleavage of DNA at specific sites by
    restriction nucleases, which greatly facilitates
    the isolation and manipulation of individual
    genes. 2. DNA cloning either through the use of
    cloning vectors or the polymerase chain reaction,
    whereby a single DNA molecule can be copied to
    generate many billions of identical molecules.
    3. Nucleic acid hybridization, which makes it
    possible to find a specific sequence of DNA or
    RNA with great accuracy and sensitivity on the
    basis of its ability to bind a complementary
    nucleic acid sequence. 4. Rapid sequencing of
    all the nucleotides in a purified DNA fragment,
    which makes it possible to identify genes and to
    deduce the amino acid sequence of the proteins
    they encode. 5. Simultaneous monitoring of the
    expression level of each gene in a cell, using
    nucleic acid microarrays that allow tens of
    thousands of hybridization reactions to be
    performed simultaneously.

23
Restriction nucleases
24
Restriction nucleases produce DNA fragments that
can be easily joined together
25
DNA Gel electrophoresis
26
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27
DNA hydridization
28
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29
Gel transferred hybridization
30
In situ hydridization
31
In situ hybridization for RNA localization
32
DNA recombination
33
DNA cloning
34
YAC
35
Human genome library
36
The synthesis of cDNA
37
cDNA clones and genomic DNA clones
38
PCR
39
PCR based gene cloning
40
PCR used in forensic science
41
Protein expression systemvia a plasmid
expression vector
42
Molecular biology research
43
Studying gene functions
  • Genetic screening
  • Monitoring gene expression
  • Site-directed mutagenesis
  • Gene replacement or knockout

44
LDA
45
Reporter gene
46
Site-directed Mutagenesis
47
Genome manipulation
48
Anti-sense RNA strategy
49
Making collections of mutant organisms
50
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51
Mouse with an engineered defect in fibroblast
growth factor 5 (FGF5)
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