Page 1
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Lesson developed under NME-ICT
Course: Botany
Subject: Plant Biotechnology
National Coordinator: Prof. S.C. Bhatla
Lesson: Cloning Vectors
Lesson Developer: Dr. Vibha G. Checker
College / Department: Botany Department, Kirori Mal College,
University of Delhi
Reviewer: Prof. Suresh Chand, Professor and Head, School of Life
Sciences, Devi Ahilya Vishwavidyalaya, Indore.
Language editor: Ms. Manisha Sharma
Department: Department of Genetics
Editor: Dr Rama Sisodia
Page 2
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Lesson developed under NME-ICT
Course: Botany
Subject: Plant Biotechnology
National Coordinator: Prof. S.C. Bhatla
Lesson: Cloning Vectors
Lesson Developer: Dr. Vibha G. Checker
College / Department: Botany Department, Kirori Mal College,
University of Delhi
Reviewer: Prof. Suresh Chand, Professor and Head, School of Life
Sciences, Devi Ahilya Vishwavidyalaya, Indore.
Language editor: Ms. Manisha Sharma
Department: Department of Genetics
Editor: Dr Rama Sisodia
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Table of Contents
Chapter – Cloning Vectors
• Introduction
? Plasmids as Cloning Vectors
? Purpose build plasmid vectors: pBR322 and pUC
series
o pBR322
o pUC series
? Ti Plasmid
? Bacterial Artificial Chromosomes (BACs)
• Bacteriophages as vectors
? Lambda ( ?) Phage
o ? Insertion vectors
o ? Replacement or Substitution vectors
? M13 Phage
• Phagemids/ Phasmids
• Cosmids
• Shuttle vectors
• Yeast artificial chromosome
• P1 artificial chromosome
• Mouse artificial chromosome
• Human artificial chromosome
• Expression vectors
• Summary
• Exercise
• Glossary
• References
• Links for animations
Page 3
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Lesson developed under NME-ICT
Course: Botany
Subject: Plant Biotechnology
National Coordinator: Prof. S.C. Bhatla
Lesson: Cloning Vectors
Lesson Developer: Dr. Vibha G. Checker
College / Department: Botany Department, Kirori Mal College,
University of Delhi
Reviewer: Prof. Suresh Chand, Professor and Head, School of Life
Sciences, Devi Ahilya Vishwavidyalaya, Indore.
Language editor: Ms. Manisha Sharma
Department: Department of Genetics
Editor: Dr Rama Sisodia
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Table of Contents
Chapter – Cloning Vectors
• Introduction
? Plasmids as Cloning Vectors
? Purpose build plasmid vectors: pBR322 and pUC
series
o pBR322
o pUC series
? Ti Plasmid
? Bacterial Artificial Chromosomes (BACs)
• Bacteriophages as vectors
? Lambda ( ?) Phage
o ? Insertion vectors
o ? Replacement or Substitution vectors
? M13 Phage
• Phagemids/ Phasmids
• Cosmids
• Shuttle vectors
• Yeast artificial chromosome
• P1 artificial chromosome
• Mouse artificial chromosome
• Human artificial chromosome
• Expression vectors
• Summary
• Exercise
• Glossary
• References
• Links for animations
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Introduction
Gene manipulation has revolutionized research. ‘Vector’in essence refers to a delivery
agent/vehicle/carrier. Cloning vector is a small molecule of DNA that can carry foreign DNA
into ahost cell and is capable of self-replication.
Source:http://genomicscience.energy.gov.
Different types of cloning vectors are available, each vector designed to perform a specific
function.
Choice of the right cloning vector is an important aspect of cloning. There are certain
essential properties which a cloning vector must possess:
Page 4
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Lesson developed under NME-ICT
Course: Botany
Subject: Plant Biotechnology
National Coordinator: Prof. S.C. Bhatla
Lesson: Cloning Vectors
Lesson Developer: Dr. Vibha G. Checker
College / Department: Botany Department, Kirori Mal College,
University of Delhi
Reviewer: Prof. Suresh Chand, Professor and Head, School of Life
Sciences, Devi Ahilya Vishwavidyalaya, Indore.
Language editor: Ms. Manisha Sharma
Department: Department of Genetics
Editor: Dr Rama Sisodia
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Table of Contents
Chapter – Cloning Vectors
• Introduction
? Plasmids as Cloning Vectors
? Purpose build plasmid vectors: pBR322 and pUC
series
o pBR322
o pUC series
? Ti Plasmid
? Bacterial Artificial Chromosomes (BACs)
• Bacteriophages as vectors
? Lambda ( ?) Phage
o ? Insertion vectors
o ? Replacement or Substitution vectors
? M13 Phage
• Phagemids/ Phasmids
• Cosmids
• Shuttle vectors
• Yeast artificial chromosome
• P1 artificial chromosome
• Mouse artificial chromosome
• Human artificial chromosome
• Expression vectors
• Summary
• Exercise
• Glossary
• References
• Links for animations
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Introduction
Gene manipulation has revolutionized research. ‘Vector’in essence refers to a delivery
agent/vehicle/carrier. Cloning vector is a small molecule of DNA that can carry foreign DNA
into ahost cell and is capable of self-replication.
Source:http://genomicscience.energy.gov.
Different types of cloning vectors are available, each vector designed to perform a specific
function.
Choice of the right cloning vector is an important aspect of cloning. There are certain
essential properties which a cloning vector must possess:
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
1. It should be able to replicate itself and the DNA segment it carries independently.
2. It should be of small size, so that it is easy to manipulate.
3. It should be easy to recover from the host cell.
4. It should be easily transferred from one cell to another by simple methods.
5. It should contain several unique restriction enzyme sites. These sites are present
only once and clustered in one locationand are known as multiple cloning sites or
polylinker. Restriction enzyme site is cleaved with a restriction enzyme to open the
cloning vector without disrupting any other region. Insert DNA fragment is digested
with the same enzyme and ligated with the vector.
6. There should be easy ways to detect their presence in host organisms. Bacterial
cloning plasmids carry an antibiotic resistance gene to distinguish the host cells that
carry vectors from the host cells that do not contain vectors.
7. There should be easy ways to detect the presence of inserted DNA.
The first recombinant DNA
Video:Stanley Cohen and Herbert Boyer's historic experiment used techniques to cut and
paste DNA to create the first custom-made organism containing recombined or
"recombinant" DNA.
Source:http://www.dnalc.org/view/15915-The-first-recombinant-DNA.html
Page 5
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Lesson developed under NME-ICT
Course: Botany
Subject: Plant Biotechnology
National Coordinator: Prof. S.C. Bhatla
Lesson: Cloning Vectors
Lesson Developer: Dr. Vibha G. Checker
College / Department: Botany Department, Kirori Mal College,
University of Delhi
Reviewer: Prof. Suresh Chand, Professor and Head, School of Life
Sciences, Devi Ahilya Vishwavidyalaya, Indore.
Language editor: Ms. Manisha Sharma
Department: Department of Genetics
Editor: Dr Rama Sisodia
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Table of Contents
Chapter – Cloning Vectors
• Introduction
? Plasmids as Cloning Vectors
? Purpose build plasmid vectors: pBR322 and pUC
series
o pBR322
o pUC series
? Ti Plasmid
? Bacterial Artificial Chromosomes (BACs)
• Bacteriophages as vectors
? Lambda ( ?) Phage
o ? Insertion vectors
o ? Replacement or Substitution vectors
? M13 Phage
• Phagemids/ Phasmids
• Cosmids
• Shuttle vectors
• Yeast artificial chromosome
• P1 artificial chromosome
• Mouse artificial chromosome
• Human artificial chromosome
• Expression vectors
• Summary
• Exercise
• Glossary
• References
• Links for animations
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Introduction
Gene manipulation has revolutionized research. ‘Vector’in essence refers to a delivery
agent/vehicle/carrier. Cloning vector is a small molecule of DNA that can carry foreign DNA
into ahost cell and is capable of self-replication.
Source:http://genomicscience.energy.gov.
Different types of cloning vectors are available, each vector designed to perform a specific
function.
Choice of the right cloning vector is an important aspect of cloning. There are certain
essential properties which a cloning vector must possess:
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
1. It should be able to replicate itself and the DNA segment it carries independently.
2. It should be of small size, so that it is easy to manipulate.
3. It should be easy to recover from the host cell.
4. It should be easily transferred from one cell to another by simple methods.
5. It should contain several unique restriction enzyme sites. These sites are present
only once and clustered in one locationand are known as multiple cloning sites or
polylinker. Restriction enzyme site is cleaved with a restriction enzyme to open the
cloning vector without disrupting any other region. Insert DNA fragment is digested
with the same enzyme and ligated with the vector.
6. There should be easy ways to detect their presence in host organisms. Bacterial
cloning plasmids carry an antibiotic resistance gene to distinguish the host cells that
carry vectors from the host cells that do not contain vectors.
7. There should be easy ways to detect the presence of inserted DNA.
The first recombinant DNA
Video:Stanley Cohen and Herbert Boyer's historic experiment used techniques to cut and
paste DNA to create the first custom-made organism containing recombined or
"recombinant" DNA.
Source:http://www.dnalc.org/view/15915-The-first-recombinant-DNA.html
Cloning Vectors
Institute of Lifelong Learning, University of Delhi
Plasmids as Cloning Vectors
Animation: Plasmids as cloning vector
Source:http://www.dnalc.org/view/15476-Mechanism-of-Recombination-3D-
animation-with-with-basic-narration.html
Source:http://upload.wikimedia.org/wikipedia/commons/thumb/c/cf/Plasmid_%28english%
29.svg/1280px-Plasmid_%28english%29.svg.png
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