shuttle vectors ppts.pptx

Shuttle vector
Made BY ALLAH NAWAZ
From uog
Vectors
 DNA molecule originating from a virus, a plasmid, or the cell of a
higher organism into which another DNA fragment of appropriate size
can be integrated without the loss of the vector capacity for self-
replication.
 Vectors introduce foreign DNA into the host cells, where it can be
reproduced in large quantities.
Shuttle vector definition
 A shuttle vector is a vector (usually a plasmid)
constructed so that it can propagate in two
different host species. It include plasmids that
can propagate in eukaryotes and prokaryotes or
in different species of bacteria
REASON FOR DEVELOPING SHUTTLE
VECTOR
 Prokaryotic vectors cannot exist & work in eukaryotic
cells because the system of two groups of organisms
varies.
 Therefore, vectors with two origin of replication were
constructed which may exist in both eukaryotes and
prokaryotes.
Shuttle vector
 Since these vectors can be grown in one host and then moved into
another without any extra manipulation, they are called shuttle vectors.
 These vectors have been designed to replicate in cells of two different
species; therefore, they contain two origins of replication, one specific
for each host species, as well as those genes necessary for their
replication and not provided by the host cells.
 These vectors are created by recombinant techniques.
 Some of them can be grown in two different prokaryotic species,
usually E. coli and a eukaryotic one, e.g. yeast, plants, animals.
 YEp13 is an example of shuttle vector.
Shuttle vector
 The 2 µm plasmid is an excellent basis for a cloning
vector. It is 6 kb in size, which is ideal for a vector, and
exists in the yeast cell at a copy number of between 70
and 200.
 Replication makes use of a plasmid origin, several
enzymes provided by the host cell, and the proteins
coded by the REP1 and REP2 genes carried by the
plasmid.
Shuttle vector
 However, all is not perfectly straightforward in using the 2
µm plasmid as a cloning vector. First, there is the question of
a selectable marker.
 In order to use LEU2 as a selectable marker, a special kind
of host organism is needed.
Shuttle vector
 Selection is possible because transformants contain a plasmid-
borne copy of the LEU2 gene, and grow in the absence of the
amino acid.
 In a cloning experiment, cells are plated out onto minimal
medium, which contains no added amino acids. Only
transformed cells are able to survive and form colonies.
shuttle vectors ppts.pptx
USES OF SHUTTLE VECTOR
 It might be difficult to recover the recombinant DNA
molecule from a transformed yeast colony.
USES OF SHUTTLE VECTOR
 This is not such a problem with YEps, which are present in yeast
cells primarily as plasmids, but with other yeast vectors, which
may integrate into one of the yeast chromosomes, purification
might be impossible.
 This is a disadvantage because in many cloning experiments
purification of recombinant DNA is essential in order for the
correct construct to be identified by, for example, DNA
sequencing.
 The standard procedure when cloning in yeast is therefore to
perform the initial cloning experiment with E. coli, and to select
recombinants in this organism.
Expression Vector definition
 The expression vectors are vectors which act as
vehicles for DNA insert and also allow the DNA insert
to be expressed efficiently. These may be plasmids or
viruses. The expression vectors are also known as
expression constructs.
shuttle vectors ppts.pptx
Expression Vector
 The expression vectors are genetically engineered for the introduction of
genes into the target cells. In addition to the gene of interest, these
expression constructs also contain regulatory elements like enhancers and
promoters so that efficient transcription of the gene of interest occurs.
 The simplest expression constructs are also known as transcription
vectors; only because they allow transcription of the cloned foreign gene
and not its translation. The vectors which facilitate both transcription and
translation of the cloned foreign gene are known as protein expression
vectors. These protein expression constructs also lead to the production of
recombinant protein.
How do Expression Vectors work?
 Once the expression construct is inside the host cell, the
protein encoded by the gene of interest is produced by
the transcription. Thereafter, it utilizes the translation
machinery and ribosomal complexes of the host
organism.
 Frequently, the plasmid is genetically engineered to
harbor regulatory elements like enhancers and
promoters. These regulator sequences aid in efficient
transcription of the gene of interest.
How do Expression Vectors work?
 Expression vectors are extensively used as tools which
help in the production of mRNAs and, in turn, stable
proteins. They are of much interest in biotechnology and
molecular biology for the production of proteins like
insulin. Insulin is the chief ingredient in the treatment of
the complex disease, Diabetes.
 When the protein product is expressed, it is to be then
purified. The purification of a protein poses a challenge
since the protein of interest, whose gene is carried on
the expression vector, is to be purified independently of
the proteins of the host organism.
Expression Vectors
 Therefore, expression vectors must have the following
expression signals:
 Strong promoter
 Strong termination codon
 Adjustment of distance between the promoter and
cloned gene
 Inserted transcription termination sequence
shuttle vectors ppts.pptx
Uses
 Expression vectors are the basic tools in
biotechnology for the production of proteins.
Escherichia coli is commonly used as the host for
protein production, but other cell types may also
be used. An example of the use of expression
vector is the production of insulin, which is used
for medical treatments of diabetes.
Differences shuttle and expression
vectors
Expression vectors are used for molecular
biology techniques such as site-directed
mutagenesis.
 A shuttle vector is a vector that can
propagate in two different host species,
hence, inserted DNA can be tested or
manipulated in two different cell types
shuttle vectors ppts.pptx
1 de 21

Recomendados

Vector Engineering.pptx por
Vector Engineering.pptxVector Engineering.pptx
Vector Engineering.pptxRenukaVyawahare
318 vistas36 diapositivas
Nucleic Acid Based Therapeutic Delivery System por
Nucleic Acid Based Therapeutic Delivery SystemNucleic Acid Based Therapeutic Delivery System
Nucleic Acid Based Therapeutic Delivery SystemKushal Saha
1.4K vistas23 diapositivas
Shuttle vector - a plasmid vector used in rDNA technology. por
Shuttle vector - a plasmid vector used in rDNA technology. Shuttle vector - a plasmid vector used in rDNA technology.
Shuttle vector - a plasmid vector used in rDNA technology. neeru02
37.8K vistas16 diapositivas
Cloning vectors & gene constructs por
Cloning vectors & gene constructsCloning vectors & gene constructs
Cloning vectors & gene constructsKavya Kondaka
4.3K vistas42 diapositivas
Gene expression vector by tahura mariyam ansari por
Gene expression vector by tahura mariyam ansariGene expression vector by tahura mariyam ansari
Gene expression vector by tahura mariyam ansariTahura Mariyam Ansari
119 vistas33 diapositivas
Expression vector por
Expression vectorExpression vector
Expression vectorAhmed Madni
6.4K vistas6 diapositivas

Más contenido relacionado

Similar a shuttle vectors ppts.pptx

Vector isolation por
Vector isolationVector isolation
Vector isolationSyedaKumail
308 vistas32 diapositivas
Cloning in eukaryotes por
Cloning in eukaryotesCloning in eukaryotes
Cloning in eukaryotesPriyengha R.S
12.5K vistas48 diapositivas
Dna cloning por
Dna cloningDna cloning
Dna cloningastro60
695 vistas21 diapositivas
DNA CLONING por
DNA CLONINGDNA CLONING
DNA CLONINGastro60
622 vistas21 diapositivas
Dna cloning por
Dna cloningDna cloning
Dna cloningastro60
6.3K vistas21 diapositivas

Similar a shuttle vectors ppts.pptx(20)

Vector isolation por SyedaKumail
Vector isolationVector isolation
Vector isolation
SyedaKumail308 vistas
Cloning in eukaryotes por Priyengha R.S
Cloning in eukaryotesCloning in eukaryotes
Cloning in eukaryotes
Priyengha R.S12.5K vistas
Dna cloning por astro60
Dna cloningDna cloning
Dna cloning
astro60695 vistas
DNA CLONING por astro60
DNA CLONINGDNA CLONING
DNA CLONING
astro60622 vistas
Dna cloning por astro60
Dna cloningDna cloning
Dna cloning
astro606.3K vistas
Vectors Used for Gene Cloning in Plants por Arunodaya Maji
Vectors Used for Gene Cloning in PlantsVectors Used for Gene Cloning in Plants
Vectors Used for Gene Cloning in Plants
Arunodaya Maji896 vistas
Microbial Genetics: Transformation, Transduction, Conjugation, Plasmids, Tran... por Theabhi.in
Microbial Genetics: Transformation, Transduction, Conjugation, Plasmids, Tran...Microbial Genetics: Transformation, Transduction, Conjugation, Plasmids, Tran...
Microbial Genetics: Transformation, Transduction, Conjugation, Plasmids, Tran...
Theabhi.in16.5K vistas
PLASMIDS AND VECTORS por Vidyashrish
PLASMIDS AND VECTORSPLASMIDS AND VECTORS
PLASMIDS AND VECTORS
Vidyashrish245 vistas
Binary_and_Shuttle_Vectors__1_.pdf por dileeptiwari28
Binary_and_Shuttle_Vectors__1_.pdfBinary_and_Shuttle_Vectors__1_.pdf
Binary_and_Shuttle_Vectors__1_.pdf
dileeptiwari2818 vistas
Vector engineering and codon optimization por Piyush Jamwal
Vector engineering and codon optimizationVector engineering and codon optimization
Vector engineering and codon optimization
Piyush Jamwal2.7K vistas
08 Kjm206 Expression Vector, Plasmid Vector por Jeneesh Jose
08 Kjm206 Expression Vector, Plasmid Vector08 Kjm206 Expression Vector, Plasmid Vector
08 Kjm206 Expression Vector, Plasmid Vector
Jeneesh Jose7.7K vistas
Transfection in animals & plants por SunandaArya
Transfection  in animals & plantsTransfection  in animals & plants
Transfection in animals & plants
SunandaArya1.8K vistas
Viral vectors in virology por Juliet Abisha
Viral vectors in virologyViral vectors in virology
Viral vectors in virology
Juliet Abisha21.3K vistas

Último

Note on the Riemann Hypothesis por
Note on the Riemann HypothesisNote on the Riemann Hypothesis
Note on the Riemann Hypothesisvegafrank2
9 vistas20 diapositivas
Exploring the nature and synchronicity of early cluster formation in the Larg... por
Exploring the nature and synchronicity of early cluster formation in the Larg...Exploring the nature and synchronicity of early cluster formation in the Larg...
Exploring the nature and synchronicity of early cluster formation in the Larg...Sérgio Sacani
1.5K vistas12 diapositivas
Worldviews and their (im)plausibility: Science and Holism por
Worldviews and their (im)plausibility: Science and HolismWorldviews and their (im)plausibility: Science and Holism
Worldviews and their (im)plausibility: Science and HolismJohnWilkins48
44 vistas19 diapositivas
A giant thin stellar stream in the Coma Galaxy Cluster por
A giant thin stellar stream in the Coma Galaxy ClusterA giant thin stellar stream in the Coma Galaxy Cluster
A giant thin stellar stream in the Coma Galaxy ClusterSérgio Sacani
23 vistas14 diapositivas
ZEBRA FISH: as model organism.pptx por
ZEBRA FISH: as model organism.pptxZEBRA FISH: as model organism.pptx
ZEBRA FISH: as model organism.pptxmahimachoudhary0807
14 vistas17 diapositivas

Último(20)

Note on the Riemann Hypothesis por vegafrank2
Note on the Riemann HypothesisNote on the Riemann Hypothesis
Note on the Riemann Hypothesis
vegafrank29 vistas
Exploring the nature and synchronicity of early cluster formation in the Larg... por Sérgio Sacani
Exploring the nature and synchronicity of early cluster formation in the Larg...Exploring the nature and synchronicity of early cluster formation in the Larg...
Exploring the nature and synchronicity of early cluster formation in the Larg...
Sérgio Sacani1.5K vistas
Worldviews and their (im)plausibility: Science and Holism por JohnWilkins48
Worldviews and their (im)plausibility: Science and HolismWorldviews and their (im)plausibility: Science and Holism
Worldviews and their (im)plausibility: Science and Holism
JohnWilkins4844 vistas
A giant thin stellar stream in the Coma Galaxy Cluster por Sérgio Sacani
A giant thin stellar stream in the Coma Galaxy ClusterA giant thin stellar stream in the Coma Galaxy Cluster
A giant thin stellar stream in the Coma Galaxy Cluster
Sérgio Sacani23 vistas
XUE: Molecular Inventory in the Inner Region of an Extremely Irradiated Proto... por Sérgio Sacani
XUE: Molecular Inventory in the Inner Region of an Extremely Irradiated Proto...XUE: Molecular Inventory in the Inner Region of an Extremely Irradiated Proto...
XUE: Molecular Inventory in the Inner Region of an Extremely Irradiated Proto...
Sérgio Sacani787 vistas
Generative AI to Accelerate Discovery of Materials por Deakin University
Generative AI to Accelerate Discovery of MaterialsGenerative AI to Accelerate Discovery of Materials
Generative AI to Accelerate Discovery of Materials
Effect of Integrated Nutrient Management on Growth and Yield of Solanaceous F... por SwagatBehera9
Effect of Integrated Nutrient Management on Growth and Yield of Solanaceous F...Effect of Integrated Nutrient Management on Growth and Yield of Solanaceous F...
Effect of Integrated Nutrient Management on Growth and Yield of Solanaceous F...
SwagatBehera96 vistas
Presentation on experimental laboratory animal- Hamster por Kanika13641
Presentation on experimental laboratory animal- HamsterPresentation on experimental laboratory animal- Hamster
Presentation on experimental laboratory animal- Hamster
Kanika136416 vistas
CYTOSKELETON STRUCTURE.ppt por EstherShobhaR
CYTOSKELETON STRUCTURE.pptCYTOSKELETON STRUCTURE.ppt
CYTOSKELETON STRUCTURE.ppt
EstherShobhaR20 vistas
Ellagic Acid and Its Metabolites as Potent and Selective Allosteric Inhibitor... por Trustlife
Ellagic Acid and Its Metabolites as Potent and Selective Allosteric Inhibitor...Ellagic Acid and Its Metabolites as Potent and Selective Allosteric Inhibitor...
Ellagic Acid and Its Metabolites as Potent and Selective Allosteric Inhibitor...
Trustlife184 vistas
Best Hybrid Event Platform.pptx por Harriet Davis
Best Hybrid Event Platform.pptxBest Hybrid Event Platform.pptx
Best Hybrid Event Platform.pptx
Harriet Davis11 vistas
INTRODUCTION TO PLANT SYSTEMATICS.pptx por RASHMI M G
INTRODUCTION TO PLANT SYSTEMATICS.pptxINTRODUCTION TO PLANT SYSTEMATICS.pptx
INTRODUCTION TO PLANT SYSTEMATICS.pptx
RASHMI M G 5 vistas
Oral_Presentation_by_Fatma (2).pdf por fatmaalmrzqi
Oral_Presentation_by_Fatma (2).pdfOral_Presentation_by_Fatma (2).pdf
Oral_Presentation_by_Fatma (2).pdf
fatmaalmrzqi8 vistas
Indian council for child welfare por RenuWaghmare2
Indian council for child welfareIndian council for child welfare
Indian council for child welfare
RenuWaghmare211 vistas

shuttle vectors ppts.pptx

  • 1. Shuttle vector Made BY ALLAH NAWAZ From uog
  • 2. Vectors  DNA molecule originating from a virus, a plasmid, or the cell of a higher organism into which another DNA fragment of appropriate size can be integrated without the loss of the vector capacity for self- replication.  Vectors introduce foreign DNA into the host cells, where it can be reproduced in large quantities.
  • 3. Shuttle vector definition  A shuttle vector is a vector (usually a plasmid) constructed so that it can propagate in two different host species. It include plasmids that can propagate in eukaryotes and prokaryotes or in different species of bacteria
  • 4. REASON FOR DEVELOPING SHUTTLE VECTOR  Prokaryotic vectors cannot exist & work in eukaryotic cells because the system of two groups of organisms varies.  Therefore, vectors with two origin of replication were constructed which may exist in both eukaryotes and prokaryotes.
  • 5. Shuttle vector  Since these vectors can be grown in one host and then moved into another without any extra manipulation, they are called shuttle vectors.  These vectors have been designed to replicate in cells of two different species; therefore, they contain two origins of replication, one specific for each host species, as well as those genes necessary for their replication and not provided by the host cells.  These vectors are created by recombinant techniques.  Some of them can be grown in two different prokaryotic species, usually E. coli and a eukaryotic one, e.g. yeast, plants, animals.  YEp13 is an example of shuttle vector.
  • 6. Shuttle vector  The 2 µm plasmid is an excellent basis for a cloning vector. It is 6 kb in size, which is ideal for a vector, and exists in the yeast cell at a copy number of between 70 and 200.  Replication makes use of a plasmid origin, several enzymes provided by the host cell, and the proteins coded by the REP1 and REP2 genes carried by the plasmid.
  • 7. Shuttle vector  However, all is not perfectly straightforward in using the 2 µm plasmid as a cloning vector. First, there is the question of a selectable marker.  In order to use LEU2 as a selectable marker, a special kind of host organism is needed.
  • 8. Shuttle vector  Selection is possible because transformants contain a plasmid- borne copy of the LEU2 gene, and grow in the absence of the amino acid.  In a cloning experiment, cells are plated out onto minimal medium, which contains no added amino acids. Only transformed cells are able to survive and form colonies.
  • 10. USES OF SHUTTLE VECTOR  It might be difficult to recover the recombinant DNA molecule from a transformed yeast colony.
  • 11. USES OF SHUTTLE VECTOR  This is not such a problem with YEps, which are present in yeast cells primarily as plasmids, but with other yeast vectors, which may integrate into one of the yeast chromosomes, purification might be impossible.  This is a disadvantage because in many cloning experiments purification of recombinant DNA is essential in order for the correct construct to be identified by, for example, DNA sequencing.  The standard procedure when cloning in yeast is therefore to perform the initial cloning experiment with E. coli, and to select recombinants in this organism.
  • 12. Expression Vector definition  The expression vectors are vectors which act as vehicles for DNA insert and also allow the DNA insert to be expressed efficiently. These may be plasmids or viruses. The expression vectors are also known as expression constructs.
  • 14. Expression Vector  The expression vectors are genetically engineered for the introduction of genes into the target cells. In addition to the gene of interest, these expression constructs also contain regulatory elements like enhancers and promoters so that efficient transcription of the gene of interest occurs.  The simplest expression constructs are also known as transcription vectors; only because they allow transcription of the cloned foreign gene and not its translation. The vectors which facilitate both transcription and translation of the cloned foreign gene are known as protein expression vectors. These protein expression constructs also lead to the production of recombinant protein.
  • 15. How do Expression Vectors work?  Once the expression construct is inside the host cell, the protein encoded by the gene of interest is produced by the transcription. Thereafter, it utilizes the translation machinery and ribosomal complexes of the host organism.  Frequently, the plasmid is genetically engineered to harbor regulatory elements like enhancers and promoters. These regulator sequences aid in efficient transcription of the gene of interest.
  • 16. How do Expression Vectors work?  Expression vectors are extensively used as tools which help in the production of mRNAs and, in turn, stable proteins. They are of much interest in biotechnology and molecular biology for the production of proteins like insulin. Insulin is the chief ingredient in the treatment of the complex disease, Diabetes.  When the protein product is expressed, it is to be then purified. The purification of a protein poses a challenge since the protein of interest, whose gene is carried on the expression vector, is to be purified independently of the proteins of the host organism.
  • 17. Expression Vectors  Therefore, expression vectors must have the following expression signals:  Strong promoter  Strong termination codon  Adjustment of distance between the promoter and cloned gene  Inserted transcription termination sequence
  • 19. Uses  Expression vectors are the basic tools in biotechnology for the production of proteins. Escherichia coli is commonly used as the host for protein production, but other cell types may also be used. An example of the use of expression vector is the production of insulin, which is used for medical treatments of diabetes.
  • 20. Differences shuttle and expression vectors Expression vectors are used for molecular biology techniques such as site-directed mutagenesis.  A shuttle vector is a vector that can propagate in two different host species, hence, inserted DNA can be tested or manipulated in two different cell types