The document presents information on recombinant DNA technology. It begins with an introduction explaining that recombinant DNA is artificially created by combining DNA sequences from different species. It then discusses some of the key tools and steps used in recombinant DNA technology, including restriction enzymes that cut DNA at specific sequences, DNA ligase that joins DNA fragments, vectors like plasmids and phages that allow DNA fragments to replicate, and suitable host cells like bacteria and yeast. The document provides examples of commonly used vectors and restriction enzymes and briefly outlines the basic process of recombinant DNA technology.
description of plasmids and types and importance of plasmids and artificial plasmids(PBR322,cosmids,phagemids) and selection of the recombinants and uses and advantages and disadvantages of the plasmids
Cloning is a technique scientists use to make exact genetic copies of living things. Genes, cells, tissues, and even whole animals can all be cloned. Some clones already exist in nature. Single-celled organisms like bacteria make exact copies of themselves each time they reproduce.
DNA cloning is the process of making multiple, identical copies of a particular piece of DNA. In a typical DNA cloning procedure, the gene or other DNA fragment of interest (perhaps a gene for a medically important human protein) is first inserted into a circular piece of DNA called a plasmid.- [https://www.khanacademy.org/science/...dna.../dna-cloning.../a/overview-dna-cloning]
Class9 DNA technology in secondary schoolssusera700ad
Biotechnology is the use of an organism, or a component of an organism or other biological system, to make a product or process.
Many forms of modern biotechnology rely on DNA technology.
DNA technology is the sequencing, analysis, and cutting-and-pasting of DNA.
Common forms of DNA technology include DNA sequencing, polymerase chain reaction, DNA cloning, and gel electrophoresis.
Biotechnology inventions can raise new practical concerns and ethical questions that must be addressed with informed input from all of society.
After the end of the presentation we’ll know -
What is cloning vector?
Why cloning vector?
History
Features of a cloning vector
Types of cloning vector
Plasmid
Bacteriophage
Cosmid
Bacterial Artificial Chromosome (BAC)
Yeast Artificial Chromosome (BAC)
Human Artificial Chromosome (HAC)
Retroviral Vectors
What determines choice of vector?
Vector in molecular gene cloning
Objectives:
After the end of the presentation we’ll know -
What is cloning vector?
Why cloning vector?
History
Features of a cloning vector
Types of cloning vector
Plasmid
Bacteriophage
Cosmid
Bacterial Artificial Chromosome (BAC)
Yeast Artificial Chromosome (BAC)
Human Artificial Chromosome (HAC)
Retroviral Vectors
What determines choice of vector?
Vector in molecular gene cloning
Cloning vector - The molecular analysis of DNA has been made possible by the cloning of DNA. The two molecules that are required for cloning are the DNA to be cloned and a cloning vector.
A cloning vector is a small piece of DNA taken from a virus, a plasmid or the cell of a higher organism, that can be stably maintained in an organism and into which a foreign DNA fragment can be inserted for cloning purposes.
Most vectors are genetically engineered.
The cloning vector is chosen according to the size and type of DNA to be cloned.
The vector therefore contains features that allow for the convenient insertion or removal of DNA fragment in or out of the vector, for example by treating the vector and the foreign DNA with a restriction enzyme and then ligating the fragments together.
After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use.
description of plasmids and types and importance of plasmids and artificial plasmids(PBR322,cosmids,phagemids) and selection of the recombinants and uses and advantages and disadvantages of the plasmids
Cloning is a technique scientists use to make exact genetic copies of living things. Genes, cells, tissues, and even whole animals can all be cloned. Some clones already exist in nature. Single-celled organisms like bacteria make exact copies of themselves each time they reproduce.
DNA cloning is the process of making multiple, identical copies of a particular piece of DNA. In a typical DNA cloning procedure, the gene or other DNA fragment of interest (perhaps a gene for a medically important human protein) is first inserted into a circular piece of DNA called a plasmid.- [https://www.khanacademy.org/science/...dna.../dna-cloning.../a/overview-dna-cloning]
Class9 DNA technology in secondary schoolssusera700ad
Biotechnology is the use of an organism, or a component of an organism or other biological system, to make a product or process.
Many forms of modern biotechnology rely on DNA technology.
DNA technology is the sequencing, analysis, and cutting-and-pasting of DNA.
Common forms of DNA technology include DNA sequencing, polymerase chain reaction, DNA cloning, and gel electrophoresis.
Biotechnology inventions can raise new practical concerns and ethical questions that must be addressed with informed input from all of society.
After the end of the presentation we’ll know -
What is cloning vector?
Why cloning vector?
History
Features of a cloning vector
Types of cloning vector
Plasmid
Bacteriophage
Cosmid
Bacterial Artificial Chromosome (BAC)
Yeast Artificial Chromosome (BAC)
Human Artificial Chromosome (HAC)
Retroviral Vectors
What determines choice of vector?
Vector in molecular gene cloning
Objectives:
After the end of the presentation we’ll know -
What is cloning vector?
Why cloning vector?
History
Features of a cloning vector
Types of cloning vector
Plasmid
Bacteriophage
Cosmid
Bacterial Artificial Chromosome (BAC)
Yeast Artificial Chromosome (BAC)
Human Artificial Chromosome (HAC)
Retroviral Vectors
What determines choice of vector?
Vector in molecular gene cloning
Cloning vector - The molecular analysis of DNA has been made possible by the cloning of DNA. The two molecules that are required for cloning are the DNA to be cloned and a cloning vector.
A cloning vector is a small piece of DNA taken from a virus, a plasmid or the cell of a higher organism, that can be stably maintained in an organism and into which a foreign DNA fragment can be inserted for cloning purposes.
Most vectors are genetically engineered.
The cloning vector is chosen according to the size and type of DNA to be cloned.
The vector therefore contains features that allow for the convenient insertion or removal of DNA fragment in or out of the vector, for example by treating the vector and the foreign DNA with a restriction enzyme and then ligating the fragments together.
After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use.
How to Create Map Views in the Odoo 17 ERPCeline George
The map views are useful for providing a geographical representation of data. They allow users to visualize and analyze the data in a more intuitive manner.
Ethnobotany and Ethnopharmacology:
Ethnobotany in herbal drug evaluation,
Impact of Ethnobotany in traditional medicine,
New development in herbals,
Bio-prospecting tools for drug discovery,
Role of Ethnopharmacology in drug evaluation,
Reverse Pharmacology.
Unit 8 - Information and Communication Technology (Paper I).pdfThiyagu K
This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
This is a presentation by Dada Robert in a Your Skill Boost masterclass organised by the Excellence Foundation for South Sudan (EFSS) on Saturday, the 25th and Sunday, the 26th of May 2024.
He discussed the concept of quality improvement, emphasizing its applicability to various aspects of life, including personal, project, and program improvements. He defined quality as doing the right thing at the right time in the right way to achieve the best possible results and discussed the concept of the "gap" between what we know and what we do, and how this gap represents the areas we need to improve. He explained the scientific approach to quality improvement, which involves systematic performance analysis, testing and learning, and implementing change ideas. He also highlighted the importance of client focus and a team approach to quality improvement.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
Palestine last event orientationfvgnh .pptxRaedMohamed3
An EFL lesson about the current events in Palestine. It is intended to be for intermediate students who wish to increase their listening skills through a short lesson in power point.
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
3. RECOMBINANT DNA
RECOMINENT DNA TECHNOLOGY
3
• Recombinant DNA is a form of artificial DNA that is created by
combining two or more sequences that would not normally occur
together through the process of gene splicing.
• Recombinant DNA Technology is a technology of joining together
of DNA molecules from two different species that are
inserted into a host organism to produce new genetic
combinations that are of value to science, medicine, agriculture and
industry.
5. RECOMINENT DNA TECHNOLOGY
5
• In conjunction with his studies of the tumor virus SV40,in
1972, Paul Berg succeeded in inserting DNA from a
bacterium into the virus' DNA. He thereby created the
first DNA molecule made of parts from different
organisms. This type of molecule became known as “
hybrid DNA" or "recombinant DNA". Among other Things,
Paul Berg’s method opened the way to creating bacteria
that produce substances used in medicines.
• Paul Berg is the "father of genetic engineering"
History of Genetic Engineering/Recombinant DNA technology
6. History of Recombinant DNA technology
RECOMINENT DNA TECHNOLOGY
6
• Boyer and Cohen's achievement represented an advance upon the
ingenious techniques developed by Paul Berg, in 1972, for inserting viral
DNA into bacterial DNA. It was a creative synthesis of earlier research that
made use of:
Living organisms able to serve as carriers for genes from another
organism.
Enzymes to cleave and rejoin DNA fragments that contain such
genes.
DNA molecules from one organism precisely targeted and
manipulated for insertion into the DNA of another organism.
8. Basic Principle of Recombinant DNA Technology
• The DNA is inserted into
another DNA molecule
called vector.
The recombinant vector is
then introduced into a
host cell where it
replicates itself and
multiple copy of gene
produced.
RECOMINENT DNA TECHNOLOGY
8
10. Recombinant DNA Technologies
RECOMINENT DNA TECHNOLOGY
10
1. Gene of interest (DNA) is isolated
(DNA fragment)
2. A desired gene is inserted into a DNA molecule – vector
(plasmid, bacteriophage or a viral genome)
3. After entering the host cell, vector grown/ replicate to form a clone.
(bacteria, yeast, plant or animal cell)
4. Large quantities of the gene product can be harvested from the clone.
13. Restriction Enzymes
RECOMINENT DNA TECHNOLOGY
13
Naturally produced by bacteria –
Restriction Endonucleases
Natural function - destroy
bacteriophage DNA in bacterial cells.
Cannot digest host DNA with
methylated C (cytosine).
14. Restriction Enzymes
RECOMINENT DNA TECHNOLOGY
14
Restriction Endonucleases (RE): Endonuclease are enzymes that produce
internal cut called cleavage in DNA molecules. A class of endonucleases that
cleaves/cut DNA only within or near those sites which have specific base
sequences, such endonucleases are known as restriction endonucleases or
restriction enzymes and site recognised by them are called recognition sequences
or recognition sites. There are three types of RE.
Type I Restriction Endonuclease
Type II Restriction Endonuclease
Type III Restriction Endonuclease
17. How RE works in Recombinant DNA Technology?
Presentation title 17
18. Ligase
RECOMINENT DNA TECHNOLOGY
18
• DNA ligase is a enzyme that can link together DNA strands that
have double-strand breaks (a break in both complementary strands
of DNA).
– Naturally DNA ligase has applications in both DNA replication
and DNA repair .
– Needs ATP
• DNA ligase has extensive use in molecular biology laboratories for
genetic recombination experiment
20. Vectors
• “A vector is a DNA molecule that has the ability to replicate
autonomously in an appropriate host cells and serve as a vehicle that
carry DNA fragment or insert to be cloned.”
• Therefore, a vector must have an origin of DNA replication (ori) that
functions in the host cell. Any extra-chromosomal small genome eg.
Plasmid, Phage or virus may be used as a vector.
21. Properties of good vector
• Able to replicate autonomously
• Small in size
• Easy to isolate and purify
• Easy transformation into host cell
• Suitable marker gene to allow easy detection and
• selection
• Multiple cloning site
• Gene Transfer: integrate DNA insert in host
• genome
• Expression: Suitable control/regulatory elements
Presentation title 21
22. Cloning vs expression vector
• Cloning vector: vector used for propagation of DNA inserts in a
suitable host are called cloning vector.
• Example: plasmid
• Expression vector: designed for expression of DNA insert i.e.
production of protein specified by inserted DNA, it is termed as
expression vector.
• Example: Bacterial cell
Presentation title 22
24. Plasmid vector
• Many different E. coli plasmids
are used as vectors.
• The natural plasmid have been
modified, shortened,
reconstructed and recombined
both in vitro and in vivo to create
plasmid of enhance utility and
specific functions.
• • Examples: pBR322,
pUC18/19, pGEM3Z, pTZ57R/T
Presentation title 24
25. Bacteriophage vectors
• Bacteriophage are viruses that attack bacteria. Several bacteriophage
are used as cloning vectors.
• The most commonly used E. coli phages are λ (lambda) and
M13phage.
• Advantage over plasmid
1. More efficient for cloning large DNA fragment (approx. 24 kb)
2. Transformation of host cell is easier with phage particle.
• Examples: λgt10, λgt11, λEMBL4, M13mp8, M13m9
Presentation title 25
26. Cosmids
• Hybrid of plasmid/Phage vectors
• contain E coli that allow it to maintained as a plasmid in the cell and carries
a λ cos site.
• A plasmid that carries a cos site.
• cos sites, act as substrates for in vitro packaging because the cos site is the
only sequence that a DNA molecule needs in order to be recognized as a 'X
genome' by the proteins that package DNA into A phage particles.
• Example: Supercosl, pJB8, pWEB, Cosmid can accommodate upto 40 kb DNA
insert
Presentation title 26
27. Continue
• Advantages: Useful for cloning very large DNA fragments (32-47 kbp).
• Disadvantages: Not easy to handle very large plasmids (~ 50 kbp).
Presentation title 27
28. Phagemid vectors
• A plasmid vector that contain origin of replication from a phage virus,
in addition to that of the plasmid is called phagemid.
• Example: pBluescript SK(+/-)
Presentation title 28
29. Phasmid vectors
• λ bacteriophage insertion vector in which their middle non essential
segment is replaced by linear plasmid with intact replication module
and lac Z gene.
• Examples: λ ZAPII, λ ZAP Express
Presentation title 29
30. Hosts for DNA Recombinant Technology
1. Bacteria
• - E. coli - used because is easily
grown and its genomics are well
• understood. Gene product is
purified from host cells
2. Yeasts - Saccharomyces
cerevisiae
• Used because it is easily grown
and its genomics are known
• May express eukaryotic genes
easily
• Easily collected and purified
3. Plant cells and whole plants
• May express eukaryotic genes
easily
• Plants are easily grown -
produce plants with new
properties.
4. Mammalian cells
• May express eukaryotic genes
easily
• Harder to grow
• Medical use.
Presentation title 30
31. Thank you
From
1. Muhammad Ramzan
2. Muhammad Kashif
3. Muhammad Reshail
4. Muhammad Sohaib
5. Muhammad Hamza
