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 von 21

Recomendados

Vector Engineering.pptx von
Vector Engineering.pptxVector Engineering.pptx
Vector Engineering.pptxRenukaVyawahare
285 views36 Folien
Nucleic Acid Based Therapeutic Delivery System von
Nucleic Acid Based Therapeutic Delivery SystemNucleic Acid Based Therapeutic Delivery System
Nucleic Acid Based Therapeutic Delivery SystemKushal Saha
1.4K views23 Folien
Shuttle vector - a plasmid vector used in rDNA technology. von
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
37.6K views16 Folien
Cloning vectors & gene constructs von
Cloning vectors & gene constructsCloning vectors & gene constructs
Cloning vectors & gene constructsKavya Kondaka
4.3K views42 Folien
Gene expression vector by tahura mariyam ansari von
Gene expression vector by tahura mariyam ansariGene expression vector by tahura mariyam ansari
Gene expression vector by tahura mariyam ansariTahura Mariyam Ansari
119 views33 Folien
Expression vector von
Expression vectorExpression vector
Expression vectorAhmed Madni
6.4K views6 Folien

Más contenido relacionado

Similar a shuttle vectors ppts.pptx

Vector isolation von
Vector isolationVector isolation
Vector isolationSyedaKumail
302 views32 Folien
Cloning in eukaryotes von
Cloning in eukaryotesCloning in eukaryotes
Cloning in eukaryotesPriyengha R.S
12.4K views48 Folien
Dna cloning von
Dna cloningDna cloning
Dna cloningastro60
695 views21 Folien
DNA CLONING von
DNA CLONINGDNA CLONING
DNA CLONINGastro60
622 views21 Folien
Dna cloning von
Dna cloningDna cloning
Dna cloningastro60
6.3K views21 Folien

Similar a shuttle vectors ppts.pptx(20)

Cloning in eukaryotes von Priyengha R.S
Cloning in eukaryotesCloning in eukaryotes
Cloning in eukaryotes
Priyengha R.S12.4K views
Dna cloning von astro60
Dna cloningDna cloning
Dna cloning
astro60695 views
DNA CLONING von astro60
DNA CLONINGDNA CLONING
DNA CLONING
astro60622 views
Dna cloning von astro60
Dna cloningDna cloning
Dna cloning
astro606.3K views
Vectors Used for Gene Cloning in Plants von Arunodaya Maji
Vectors Used for Gene Cloning in PlantsVectors Used for Gene Cloning in Plants
Vectors Used for Gene Cloning in Plants
Arunodaya Maji870 views
Microbial Genetics: Transformation, Transduction, Conjugation, Plasmids, Tran... von Theabhi.in
Microbial Genetics: Transformation, Transduction, Conjugation, Plasmids, Tran...Microbial Genetics: Transformation, Transduction, Conjugation, Plasmids, Tran...
Microbial Genetics: Transformation, Transduction, Conjugation, Plasmids, Tran...
Theabhi.in16.2K views
PLASMIDS AND VECTORS von Vidyashrish
PLASMIDS AND VECTORSPLASMIDS AND VECTORS
PLASMIDS AND VECTORS
Vidyashrish245 views
Binary_and_Shuttle_Vectors__1_.pdf von dileeptiwari28
Binary_and_Shuttle_Vectors__1_.pdfBinary_and_Shuttle_Vectors__1_.pdf
Binary_and_Shuttle_Vectors__1_.pdf
dileeptiwari2818 views
Vector engineering and codon optimization von Piyush Jamwal
Vector engineering and codon optimizationVector engineering and codon optimization
Vector engineering and codon optimization
Piyush Jamwal2.7K views
08 Kjm206 Expression Vector, Plasmid Vector von Jeneesh Jose
08 Kjm206 Expression Vector, Plasmid Vector08 Kjm206 Expression Vector, Plasmid Vector
08 Kjm206 Expression Vector, Plasmid Vector
Jeneesh Jose7.7K views
Transfection in animals & plants von SunandaArya
Transfection  in animals & plantsTransfection  in animals & plants
Transfection in animals & plants
SunandaArya1.8K views
Viral vectors in virology von Juliet Abisha
Viral vectors in virologyViral vectors in virology
Viral vectors in virology
Juliet Abisha21.3K views

Último

Workshop Chemical Robotics ChemAI 231116.pptx von
Workshop Chemical Robotics ChemAI 231116.pptxWorkshop Chemical Robotics ChemAI 231116.pptx
Workshop Chemical Robotics ChemAI 231116.pptxMarco Tibaldi
95 views41 Folien
"How can I develop my learning path in bioinformatics? von
"How can I develop my learning path in bioinformatics?"How can I develop my learning path in bioinformatics?
"How can I develop my learning path in bioinformatics?Bioinformy
18 views13 Folien
1978 NASA News Release Log von
1978 NASA News Release Log1978 NASA News Release Log
1978 NASA News Release Logpurrterminator
7 views146 Folien
A Ready-to-Analyze High-Plex Spatial Signature Development Workflow for Cance... von
A Ready-to-Analyze High-Plex Spatial Signature Development Workflow for Cance...A Ready-to-Analyze High-Plex Spatial Signature Development Workflow for Cance...
A Ready-to-Analyze High-Plex Spatial Signature Development Workflow for Cance...InsideScientific
9 views62 Folien
SANJAY HPLC.pptx von
SANJAY HPLC.pptxSANJAY HPLC.pptx
SANJAY HPLC.pptxsanjayudps2016
142 views38 Folien
Ecology von
Ecology Ecology
Ecology Abhijith Raj.R
6 views10 Folien

Último(20)

Workshop Chemical Robotics ChemAI 231116.pptx von Marco Tibaldi
Workshop Chemical Robotics ChemAI 231116.pptxWorkshop Chemical Robotics ChemAI 231116.pptx
Workshop Chemical Robotics ChemAI 231116.pptx
Marco Tibaldi95 views
"How can I develop my learning path in bioinformatics? von Bioinformy
"How can I develop my learning path in bioinformatics?"How can I develop my learning path in bioinformatics?
"How can I develop my learning path in bioinformatics?
Bioinformy18 views
A Ready-to-Analyze High-Plex Spatial Signature Development Workflow for Cance... von InsideScientific
A Ready-to-Analyze High-Plex Spatial Signature Development Workflow for Cance...A Ready-to-Analyze High-Plex Spatial Signature Development Workflow for Cance...
A Ready-to-Analyze High-Plex Spatial Signature Development Workflow for Cance...
himalay baruah acid fast staining.pptx von HimalayBaruah
himalay baruah acid fast staining.pptxhimalay baruah acid fast staining.pptx
himalay baruah acid fast staining.pptx
HimalayBaruah5 views
Open Access Publishing in Astrophysics von Peter Coles
Open Access Publishing in AstrophysicsOpen Access Publishing in Astrophysics
Open Access Publishing in Astrophysics
Peter Coles543 views
Ethical issues associated with Genetically Modified Crops and Genetically Mod... von PunithKumars6
Ethical issues associated with Genetically Modified Crops and Genetically Mod...Ethical issues associated with Genetically Modified Crops and Genetically Mod...
Ethical issues associated with Genetically Modified Crops and Genetically Mod...
PunithKumars618 views
Metatheoretical Panda-Samaneh Borji.pdf von samanehborji
Metatheoretical Panda-Samaneh Borji.pdfMetatheoretical Panda-Samaneh Borji.pdf
Metatheoretical Panda-Samaneh Borji.pdf
samanehborji16 views
Synthesis and Characterization of Magnetite-Magnesium Sulphate-Sodium Dodecyl... von GIFT KIISI NKIN
Synthesis and Characterization of Magnetite-Magnesium Sulphate-Sodium Dodecyl...Synthesis and Characterization of Magnetite-Magnesium Sulphate-Sodium Dodecyl...
Synthesis and Characterization of Magnetite-Magnesium Sulphate-Sodium Dodecyl...
GIFT KIISI NKIN14 views
Distinct distributions of elliptical and disk galaxies across the Local Super... von Sérgio Sacani
Distinct distributions of elliptical and disk galaxies across the Local Super...Distinct distributions of elliptical and disk galaxies across the Local Super...
Distinct distributions of elliptical and disk galaxies across the Local Super...
Sérgio Sacani30 views
ENTOMOLOGY PPT ON BOMBYCIDAE AND SATURNIIDAE.pptx von MN
ENTOMOLOGY PPT ON BOMBYCIDAE AND SATURNIIDAE.pptxENTOMOLOGY PPT ON BOMBYCIDAE AND SATURNIIDAE.pptx
ENTOMOLOGY PPT ON BOMBYCIDAE AND SATURNIIDAE.pptx
MN6 views
Light Pollution for LVIS students von CWBarthlmew
Light Pollution for LVIS studentsLight Pollution for LVIS students
Light Pollution for LVIS students
CWBarthlmew5 views

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