Lecture 10 - Introduction to Recombinant DNA Technology | Plant Biotechnology - Botany PDF Download

 Page 1


 Introduction to recombinant DNA technology  
1 
Institute of Lifelong Learning, University of Delhi 
                                                                                              
 
 
 
 
 
 
 
Discipline: Botany 
Paper: Plant Biotechnology 
National Coordinator: Prof. S.C. Bhatla 
Lesson: Introduction to Recombinant DNA Technology 
Lesson Developer: Ms. Manisha Sharma, UDSC 
Department/College: Genetics 
 
 
 
 
 
 
 
 
 
Page 2


 Introduction to recombinant DNA technology  
1 
Institute of Lifelong Learning, University of Delhi 
                                                                                              
 
 
 
 
 
 
 
Discipline: Botany 
Paper: Plant Biotechnology 
National Coordinator: Prof. S.C. Bhatla 
Lesson: Introduction to Recombinant DNA Technology 
Lesson Developer: Ms. Manisha Sharma, UDSC 
Department/College: Genetics 
 
 
 
 
 
 
 
 
 
 Introduction to recombinant DNA technology  
2 
Institute of Lifelong Learning, University of Delhi 
Table of Contents 
Introduction to recombinant DNA technology 
? What is recombinant DNA? 
? But first, what is DNA? 
? History – Early experiments in recombinant DNA technology 
? Applications of recombinant DNA technology 
? Pharmaceutical uses 
? Vaccines 
? Diagnostic testing 
? Gene therapy 
? Monoclonal antibodies 
? DNA fingerprinting 
? Food additives 
? Agricultural uses 
? Animal husbandry 
? Summary 
? Questions 
? Glossary 
? References 
? Web links  
Page 3


 Introduction to recombinant DNA technology  
1 
Institute of Lifelong Learning, University of Delhi 
                                                                                              
 
 
 
 
 
 
 
Discipline: Botany 
Paper: Plant Biotechnology 
National Coordinator: Prof. S.C. Bhatla 
Lesson: Introduction to Recombinant DNA Technology 
Lesson Developer: Ms. Manisha Sharma, UDSC 
Department/College: Genetics 
 
 
 
 
 
 
 
 
 
 Introduction to recombinant DNA technology  
2 
Institute of Lifelong Learning, University of Delhi 
Table of Contents 
Introduction to recombinant DNA technology 
? What is recombinant DNA? 
? But first, what is DNA? 
? History – Early experiments in recombinant DNA technology 
? Applications of recombinant DNA technology 
? Pharmaceutical uses 
? Vaccines 
? Diagnostic testing 
? Gene therapy 
? Monoclonal antibodies 
? DNA fingerprinting 
? Food additives 
? Agricultural uses 
? Animal husbandry 
? Summary 
? Questions 
? Glossary 
? References 
? Web links  
 Introduction to recombinant DNA technology  
3 
Institute of Lifelong Learning, University of Delhi 
Learning outcomes 
? After reading this article, the reader should have understood how a genetically 
modified organism is created. 
? The reader should be able to differentiate between recombinant DNA and the 
process of genetic recombination. 
? The reader should have known the history and major accomplishments which 
have led to the initiation of recombinant DNA technology. 
? The reader should be able to identify how and in what fields recombinant DNA 
technology is being used today and the major areas of focus where this 
technology is being developed. 
Introduction 
What is recombinant DNA? 
Recombinant DNA, or rDNA, as the name suggests, is comprised of two or more pieces 
of DNA molecules that have been combined together to form a single molecule. 
 
 
Figure: Diagrammatic representation of two DNA molecules that join together to form a 
recombinant DNA molecule. 
Source: Developed by author 
 
Generally, creation of a recombinant DNA molecule involves two steps: 
? In the first step, DNA from two or more sources containing a gene or any other 
fragment of interest are cut at specific points/sites. 
DNA 1 DNA 2
+
Recombinant DNA
Page 4


 Introduction to recombinant DNA technology  
1 
Institute of Lifelong Learning, University of Delhi 
                                                                                              
 
 
 
 
 
 
 
Discipline: Botany 
Paper: Plant Biotechnology 
National Coordinator: Prof. S.C. Bhatla 
Lesson: Introduction to Recombinant DNA Technology 
Lesson Developer: Ms. Manisha Sharma, UDSC 
Department/College: Genetics 
 
 
 
 
 
 
 
 
 
 Introduction to recombinant DNA technology  
2 
Institute of Lifelong Learning, University of Delhi 
Table of Contents 
Introduction to recombinant DNA technology 
? What is recombinant DNA? 
? But first, what is DNA? 
? History – Early experiments in recombinant DNA technology 
? Applications of recombinant DNA technology 
? Pharmaceutical uses 
? Vaccines 
? Diagnostic testing 
? Gene therapy 
? Monoclonal antibodies 
? DNA fingerprinting 
? Food additives 
? Agricultural uses 
? Animal husbandry 
? Summary 
? Questions 
? Glossary 
? References 
? Web links  
 Introduction to recombinant DNA technology  
3 
Institute of Lifelong Learning, University of Delhi 
Learning outcomes 
? After reading this article, the reader should have understood how a genetically 
modified organism is created. 
? The reader should be able to differentiate between recombinant DNA and the 
process of genetic recombination. 
? The reader should have known the history and major accomplishments which 
have led to the initiation of recombinant DNA technology. 
? The reader should be able to identify how and in what fields recombinant DNA 
technology is being used today and the major areas of focus where this 
technology is being developed. 
Introduction 
What is recombinant DNA? 
Recombinant DNA, or rDNA, as the name suggests, is comprised of two or more pieces 
of DNA molecules that have been combined together to form a single molecule. 
 
 
Figure: Diagrammatic representation of two DNA molecules that join together to form a 
recombinant DNA molecule. 
Source: Developed by author 
 
Generally, creation of a recombinant DNA molecule involves two steps: 
? In the first step, DNA from two or more sources containing a gene or any other 
fragment of interest are cut at specific points/sites. 
DNA 1 DNA 2
+
Recombinant DNA
 Introduction to recombinant DNA technology  
4 
Institute of Lifelong Learning, University of Delhi 
? In the second step, the cut DNA sequences are joined together in a controlled 
manner to create a recombinant DNA molecule. 
 
Recombinant DNA molecules are also known as chimeric DNA, because they are hybrid 
DNA molecules which have been engineered from at least two different sources, like the 
mythical chimera. 
An organism, if its genome has been altered using recombinant DNA technology is 
known as a transgenic or genetically modified (GM) organism. A transgenic organism is 
created either by combining DNA from different genomes or by inserting foreign DNA 
into the host organism?s genome. The transgene should be introgressed into the germ 
line of the organism and is stably inherited by its progeny. 
Proteins that are produced in a genetically modified organism due to the expression of a 
recombinant DNA are known as recombinant proteins. 
But first, what is DNA? 
? DNA (deoxyribonucleic acid) is the repository of genetic information that we 
inherit from our parents. It decides how we turn out to be as an organism, e.g. 
the colour of our skin, our height, curly or straight hair, even in plants whether 
they have rounded or pointed leaves, etc. DNA is involved in the creation, 
development as well as maintenance of normal body function of all organisms. 
? All organisms except some viruses have DNA as their genetic material. 
? DNA is a nucleic acid made up of long strands of sugar, phosphate and 
nitrogenous bases. 
? The nitrogenous bases can further be of four types – adenine (A), guanine (G), 
thymine (T) or cytosine(C). Adenine and guanine are purines and thymine and 
cytosine are pyrimidines. 
A gene is the basic physical and functional unit of heredity 
in any living organism. It is the coding sequence of DNA 
that contains information to make proteins in a cell.  
Page 5


 Introduction to recombinant DNA technology  
1 
Institute of Lifelong Learning, University of Delhi 
                                                                                              
 
 
 
 
 
 
 
Discipline: Botany 
Paper: Plant Biotechnology 
National Coordinator: Prof. S.C. Bhatla 
Lesson: Introduction to Recombinant DNA Technology 
Lesson Developer: Ms. Manisha Sharma, UDSC 
Department/College: Genetics 
 
 
 
 
 
 
 
 
 
 Introduction to recombinant DNA technology  
2 
Institute of Lifelong Learning, University of Delhi 
Table of Contents 
Introduction to recombinant DNA technology 
? What is recombinant DNA? 
? But first, what is DNA? 
? History – Early experiments in recombinant DNA technology 
? Applications of recombinant DNA technology 
? Pharmaceutical uses 
? Vaccines 
? Diagnostic testing 
? Gene therapy 
? Monoclonal antibodies 
? DNA fingerprinting 
? Food additives 
? Agricultural uses 
? Animal husbandry 
? Summary 
? Questions 
? Glossary 
? References 
? Web links  
 Introduction to recombinant DNA technology  
3 
Institute of Lifelong Learning, University of Delhi 
Learning outcomes 
? After reading this article, the reader should have understood how a genetically 
modified organism is created. 
? The reader should be able to differentiate between recombinant DNA and the 
process of genetic recombination. 
? The reader should have known the history and major accomplishments which 
have led to the initiation of recombinant DNA technology. 
? The reader should be able to identify how and in what fields recombinant DNA 
technology is being used today and the major areas of focus where this 
technology is being developed. 
Introduction 
What is recombinant DNA? 
Recombinant DNA, or rDNA, as the name suggests, is comprised of two or more pieces 
of DNA molecules that have been combined together to form a single molecule. 
 
 
Figure: Diagrammatic representation of two DNA molecules that join together to form a 
recombinant DNA molecule. 
Source: Developed by author 
 
Generally, creation of a recombinant DNA molecule involves two steps: 
? In the first step, DNA from two or more sources containing a gene or any other 
fragment of interest are cut at specific points/sites. 
DNA 1 DNA 2
+
Recombinant DNA
 Introduction to recombinant DNA technology  
4 
Institute of Lifelong Learning, University of Delhi 
? In the second step, the cut DNA sequences are joined together in a controlled 
manner to create a recombinant DNA molecule. 
 
Recombinant DNA molecules are also known as chimeric DNA, because they are hybrid 
DNA molecules which have been engineered from at least two different sources, like the 
mythical chimera. 
An organism, if its genome has been altered using recombinant DNA technology is 
known as a transgenic or genetically modified (GM) organism. A transgenic organism is 
created either by combining DNA from different genomes or by inserting foreign DNA 
into the host organism?s genome. The transgene should be introgressed into the germ 
line of the organism and is stably inherited by its progeny. 
Proteins that are produced in a genetically modified organism due to the expression of a 
recombinant DNA are known as recombinant proteins. 
But first, what is DNA? 
? DNA (deoxyribonucleic acid) is the repository of genetic information that we 
inherit from our parents. It decides how we turn out to be as an organism, e.g. 
the colour of our skin, our height, curly or straight hair, even in plants whether 
they have rounded or pointed leaves, etc. DNA is involved in the creation, 
development as well as maintenance of normal body function of all organisms. 
? All organisms except some viruses have DNA as their genetic material. 
? DNA is a nucleic acid made up of long strands of sugar, phosphate and 
nitrogenous bases. 
? The nitrogenous bases can further be of four types – adenine (A), guanine (G), 
thymine (T) or cytosine(C). Adenine and guanine are purines and thymine and 
cytosine are pyrimidines. 
A gene is the basic physical and functional unit of heredity 
in any living organism. It is the coding sequence of DNA 
that contains information to make proteins in a cell.  
 Introduction to recombinant DNA technology  
5 
Institute of Lifelong Learning, University of Delhi 
 
Figure: DNA is composed of sugar, phosphate and nitrogenous bases. Two DNA strands 
are connected by hydrogen bonds between the bases. 
Source: http://www2.le.ac.uk/departments/genetics/vgec/schoolscolleges/topics/dna-
genes-chromosomes 
 
? Nitrogenous bases on two separate DNA strands interact with each other to form 
two kinds of hydrogen bonds or H-bonds, i.e., a double bond between bases A 
and T and a triple bond between bases G and C. The two strands form a helical 
ladder-like structure, known as the DNA „double helix?. 
 
 
 
 
The three dimensional DNA double helix structure was 
proposed in 1953 by American biologist James Watson and 
English physicist Francis Crick. Their proposition was based 
on the reasoning given by Austrian biochemist Erwin 
Chargaff that A = T and C = G and the crucial X-ray 
crystallographic evidence provided by English researchers 
Rosalind Franklin and Maurice Wilkins.