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                     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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FAQs on Lecture 12 - Cloning Vectors - Plant Biotechnology - Botany

1. What are cloning vectors in botany?
Ans. Cloning vectors in botany are small DNA molecules that are used to carry and replicate foreign DNA fragments in the laboratory. These vectors are often derived from plasmids or viruses and are used to transfer genes of interest into host plant cells.
2. How are cloning vectors used in botany research?
Ans. Cloning vectors are essential tools in botany research as they allow scientists to insert and replicate specific DNA fragments in plant cells. By using these vectors, researchers can study the function of genes, engineer plants with desired traits, and explore plant genetic diversity.
3. What are the advantages of using cloning vectors in botany?
Ans. Some advantages of using cloning vectors in botany research include their ability to transfer genes between different plant species, their high replication efficiency, and their versatility in accommodating various sizes of DNA fragments. Cloning vectors also allow for the stable maintenance and expression of the inserted genes in plant cells.
4. What are the common types of cloning vectors used in botany?
Ans. The most common types of cloning vectors used in botany include plasmids, bacteriophages, and binary vectors. Plasmids are circular DNA molecules that can replicate independently in bacterial cells and can be easily manipulated in the laboratory. Bacteriophages are viruses that infect bacteria and can carry foreign DNA. Binary vectors are complex cloning vectors that are used for the transformation of plant cells.
5. How can cloning vectors contribute to crop improvement in botany?
Ans. Cloning vectors play a crucial role in crop improvement in botany by allowing for the introduction of beneficial genes into plants. These genes can confer traits such as disease resistance, drought tolerance, and increased yield. By using cloning vectors, scientists can modify the genetic makeup of crops to enhance their productivity and sustainability.
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