Chapter 6 Gene Technology

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Chapter 6Gene Technology
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Chapter review
Gene technology
Ethics
DNA recombinant technology
Gene library
Application
Genetic engineering
Gene cloning
Genomic library
cDNA library
Transgenic organism
DNA finger printing
tools
Genetically modified organism
Target gene
Cloning vector
Restriction enzyme
Modifying enzyme
Host cell
Polymerase chain reaction
methods
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At the end of the lesson, you should be able to

• Explain Recombinant DNA Technology (genetic
  engineering)
• Explain gene cloning and its application.
• List the tools used in recombinant DNA
  technology.
• Explain restriction enzyme

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About DNA

• Presents in a chromosome cell and contains
  genetic information for all living organisms.

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Genetic Engineering
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What is genetic engineering?

• The ability to take a specific gene from one cell
  and place it into another cell where it is
  expressed _at_ the direct manipulation of genes for
  practical purposes.

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What is genetic engineering?
What is genetic engineering?

• New DNA made by combining different DNA pieces
  is known as recombinant DNA.

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Biotechnology

• The manipulation of living organisms of their
  components to produce useful products.

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Biotechnology

• Used various techniques to changed living
  organisms at the molecular level to produce/
  provide good product and services.
• Often referred to as genetic engineering.

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Recombinant DNA Technology
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Recombinant DNA Technology

• Special techniques produced from genetic
  engineering process.
• Technology that produced any DNA molecule made
  in vitro with segments from different
  sources/by combining genes from different
  organisms.

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Recombinant DNA Technology

• A new field of biology - started in the mid
  1970s
• Permits the formation of new combinations of
  genes
• By isolating genes from the organisms
• And introducing them into either a similar or
  unrelated organism.

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Recombinant DNA Technology

• By this method
• foreign DNA may be inserted not only in the
  simple single cells of bacteria but also into
  cells from the bodies of complex organisms
• once inside the cells, this foreign DNA may be
  expressed.

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Recombinant DNA Technology Gene Cloning
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Gene cloning

• DNA cloning is using the DNA manipulation
  procedures to produce multiple copies of a single
  gene or segment of DNA.

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Gene cloning
Stages in gene cloning
Gene preparation
Vector insertion
Detection of cloned gene
Host cell transformation
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Gene cloning
Stages in gene cloning
Gene preparation
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Gene cloning
Stages in gene cloning
Vector insertion
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Gene cloning
Stages in gene cloning
Host cell transformation
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Recombinant DNA Technology Tools
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Tools
DNA (gene) source
DNA cloning vector
DNA ligase
Restriction enzyme
Host cell
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Chapter review
Gene technology
Types
YAC
Polymerase chain reaction
Target gene
bacteriophage
Taq Polymerase
cosmid
plasmid
Cloning vector
DNA ligase
Tools of DNA Recombinant Technology
Modifying enzyme
Types
characteristics
Host cell
Restriction enzyme
E. coli
characteristic
Types
Palindromic sequence concept
EcoRI
SmaI
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Restriction enzymes

• Enzymes for cutting DNA

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Restriction enzymes

• Class of enzyme that recognizes specific base
  sequences of DNA and cut / cleaves the DNA
  molecular at that side.
• Today, more than 800 different types known.

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Restriction enzymes

• In natural, these enzymes protect the bacteria
  against intruding (interfering) DNA/RNA from
  another organism like viruses.
• The bacterial DNA was modified, so that it was
  immuned to its own restriction enzyme

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Restriction enzymes
Characteristics

• Have ability to splice through DNA at specific
  sequence - different restriction enzyme cut a
  particular base sequences, called restriction
  sites.

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Restriction enzymes
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Restriction enzymes
Characteristics

• This enzymes recognize palindromic base sequences.

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Restriction enzymes
Characteristics
palindrome

• The base sequences of one strand reads the same
  as its compliment strands

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Restriction enzymes
Characteristics
palindrome
SPELL THIS!
M
A
D
A
M
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Restriction enzymes
Characteristics

• Example

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G A A T T C
C T T A A G
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3
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Restriction enzymes
Characteristics

• RE hydrolyze the phophodiester backbone of each
  strand

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Restriction enzymes
Characteristics

• RE can produce two types of cutting pattern
• sticky end (EcoRI)
• blunt end (SmaI)

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Restriction enzymes
Characteristics

EcoRI cutting pattern
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Restriction enzymes
Characteristics
SmaI cutting pattern
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Several types of restriction enymes
Types of RE