Chapter review…Chapter review…Gene technologyGene technology
DNA DNA recombinant recombinant technologytechnology
Genetic Genetic engineerinengineerin
gg
Gene Gene cloningcloning
toolstools
Target Target genegene
Cloning Cloning vectorvector
RestrictioRestriction enzymen enzyme
Modifying Modifying enzymeenzyme
Host cellHost cell Polymerase Polymerase chain chain
reactionreaction
methodsmethods
Gene Gene librarylibrary
Genomic Genomic librarylibrary
cDNA cDNA librarlibrar
yy
Application Application
TransgenTransgenic ic
organismorganismGenetically Genetically modified modified organismorganism
DNA DNA finger finger
printingprinting
EthicsEthics
Explain Recombinant DNA Technology (genetic engineering)
Explain gene cloning and its application.
List the tools used in recombinant DNA technology.
Explain restriction enzyme
Presents in a Presents in a chromosome cell chromosome cell and contains and contains genetic genetic information for all information for all living organisms.living organisms.
The ability to The ability to take a specific take a specific genegene from one cell and place from one cell and place it into another cell where it it into another cell where it
is expressed @ is expressed @ the direct the direct manipulation of genes for manipulation of genes for
practical purposes.practical purposes.
New DNA New DNA made by made by
combining combining different DNA different DNA
piecespieces is is known as known as
recombinant recombinant DNA.DNA.
The manipulation of living organisms of their components to produce
useful products.
Used Used various techniquesvarious techniques to changed living to changed living organisms at the molecular organisms at the molecular level to produce/ provide level to produce/ provide good product and services. good product and services.
Often referred to as Often referred to as genetic engineering.genetic engineering.
Special techniques produced Special techniques produced from genetic engineering from genetic engineering process.process.
Technology that produced any Technology that produced any DNA molecule made ‘in vitro’ DNA molecule made ‘in vitro’ with segments from different with segments from different sources/by combining genes sources/by combining genes from different organisms.from different organisms.
A new field of biology - started A new field of biology - started in the mid 1970’s in the mid 1970’s
Permits the formation of new Permits the formation of new combinations of genescombinations of genes
By isolating genes from the By isolating genes from the organismsorganisms
And introducing them into And introducing them into either a similar or unrelated either a similar or unrelated organism.organism.
By this methodBy this methoda.a.foreign DNA may be insertedforeign DNA may be inserted not only in the simple single not only in the simple single cells of bacteria but also into cells of bacteria but also into cells from the bodies of cells from the bodies of complex organismscomplex organisms
b.b.once inside the cells, this once inside the cells, this foreign DNA foreign DNA may be may be expressed.expressed.
DNA cloning is DNA cloning is using the using the DNA manipulation DNA manipulation proceduresprocedures to produce to produce multiple multiple copiescopies of a single of a single genegene or segment of DNA. or segment of DNA.
Stages in gene cloningStages in gene cloning
Gene Gene preparationpreparation Vector Vector
insertioninsertion
Host cell Host cell transformationtransformation
Detection Detection of cloned of cloned
genegene
Stages in gene Stages in gene cloningcloning
Gene Gene preparationpreparation
Stages in gene Stages in gene cloningcloning
Vector Vector insertioninsertion
Stages in gene Stages in gene cloningcloning
Host cell Host cell transformationtransformation
DNA DNA (gene) (gene) sourcesource
DNA DNA cloning cloning vectorvector
Restriction Restriction enzymeenzyme
DNA DNA ligaseligase
Host Host cellcell
Tools of DNA Tools of DNA Recombinant Recombinant TechnologyTechnology
Target Target genegene
Cloning Cloning vectorvector
RestrictioRestriction enzymen enzyme
Modifying Modifying enzymeenzyme
Host cellHost cell
Polymerase Polymerase chain chain reactionreaction
Chapter review…Chapter review…Gene technologyGene technology
TypesTypes
plasmiplasmidd
cosmidcosmidbacteriophagbacteriophagee
YACYAC
characteristiccharacteristicss
Types Types Palindromic Palindromic sequence sequence conceptconcept
DNA DNA ligaseligase
Taq Taq PolymeraPolymerasese
Types Types
EcoEcoRIRISmaSmaII
E. E. colicoli
characteristcharacteristicic
Enzymes Enzymes for for cutting cutting DNA…DNA…
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.
In natural, these enzymes protect In natural, these enzymes protect the bacteria against intruding the bacteria against intruding (interfering) DNA/RNA from (interfering) DNA/RNA from another organism like viruses.another organism like viruses.
The bacterial DNA was modified, The bacterial DNA was modified, so that it was immuned to its own so that it was immuned to its own restriction enzymerestriction enzyme
Have ability to splice through DNA at specific sequence - different restriction enzyme cut a particular base sequences, called restriction sites.
CharacteristicsCharacteristics
This enzymes This enzymes recognizerecognize palindromic base sequences.base sequences.
CharacteristicsCharacteristics
The base sequences of one strand reads the same as its
compliment strands
CharacteristicsCharacteristics
palindromepalindrome
CharacteristicsCharacteristics
palindromepalindromeSPELL THIS!SPELL THIS!
MM AA DD AA MM
Example :Example :
Characteristics Characteristics
G A A T T CG A A T T C
C T T A A GC T T A A G5’5’
5’5’ 3’3’
3’3’
RE hydrolyze the phophodiester backbone of each strand
Characteristics Characteristics
RE can produce two types of cutting pattern : sticky end (EcoRI) blunt end (SmaI)
Characteristics Characteristics
Characteristics Characteristics
EcoEcoRI RI cutting cutting patternpattern
Characteristics Characteristics
SmaSmaI I cutting cutting patternpattern
Types of RE