Recombinant DNA technology allows DNA from different sources to be combined to form artificial DNA molecules. This is done by cutting the DNA with restriction enzymes and joining the pieces together with DNA ligase. The artificial DNA can then be inserted into host cells where it is replicated. This technology was developed in 1973 and has many important applications, including producing human insulin in bacteria to treat diabetes, creating genetically modified crops with desirable traits, and producing other proteins and vaccines. The basic steps involve isolating DNA, cutting it with restriction enzymes, ligating the pieces, introducing the DNA into host cells, replicating the DNA within the cells, and identifying cells containing the recombinant DNA.
Gene Cloning Vectors - Plasmids, Bacteriophages and Phagemids.Ambika Prajapati
A cloning vector is a small piece of DNA that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes. The cloning vector may be DNA taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium.
They allow the exogenous DNA to be inserted, stored, and manipulated mainly at DNA level.
Types -
1.Plasmid vectors.
2.Bacteriophage vectors .
3.Phagemids.
To modifying the structure of a specific gene.
Gene targeting vector introduced into the cell.
Vector modifies the normal chromosomal gene through homologous recombination.
Useful in treating some human genetic disorders – Hemophilia, Duchenne Muscular Dystrophy.
Treating human diseases by genetic approaches – Gene Therapy.
Gene Therapy – Replacing the defective gene by normal copy of the gene.
Expressed sequence tag/EST is a short partial sequence, typically 200-400 bp long, of a complimentary DNA/Cdna.
EST is a short sub-sequence of a cDNA sequence.
Used to identify gene transcripts, and are instrumental in gene discovery and in gene-sequence determination.
Approximately 74.2 million ESTs are available in public databases.
EST results from one-short sequencing of a cloned cDNA.
Low-quality fragments.
Length is approximately 500 to 800 nucleotides.
Gene Cloning Vectors - Plasmids, Bacteriophages and Phagemids.Ambika Prajapati
A cloning vector is a small piece of DNA that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes. The cloning vector may be DNA taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium.
They allow the exogenous DNA to be inserted, stored, and manipulated mainly at DNA level.
Types -
1.Plasmid vectors.
2.Bacteriophage vectors .
3.Phagemids.
To modifying the structure of a specific gene.
Gene targeting vector introduced into the cell.
Vector modifies the normal chromosomal gene through homologous recombination.
Useful in treating some human genetic disorders – Hemophilia, Duchenne Muscular Dystrophy.
Treating human diseases by genetic approaches – Gene Therapy.
Gene Therapy – Replacing the defective gene by normal copy of the gene.
Expressed sequence tag/EST is a short partial sequence, typically 200-400 bp long, of a complimentary DNA/Cdna.
EST is a short sub-sequence of a cDNA sequence.
Used to identify gene transcripts, and are instrumental in gene discovery and in gene-sequence determination.
Approximately 74.2 million ESTs are available in public databases.
EST results from one-short sequencing of a cloned cDNA.
Low-quality fragments.
Length is approximately 500 to 800 nucleotides.
Plant transformation permits the introduction of the gene of interest for producing novel transgenic plants. "When a gene from one species is moved or relocated to another species by using recombinant DNA technology are called genetically modified organisms. Genetic engineering is one way to modify the plants by selecting for desired traits. Genetically modified organisms have foreign genes derive from not only plant source but also from bacteria, viruses, fungi, insects and animals. Transformation is the introduction and addition of the desired gene in plant for the generation of transgenic plant. Plant transformation is a challenging process for scientists. DNA transfer by artificial methods like DNA transfer through physical method is micro-injection, biolistic or gene gun methods, electroporation, silica carbide, microinjection, lipofection, microinjection. DNA also transfers by chemical methods. In natural method like in biological method, Agrobacterium-mediated transfer, Rhizobium, virus-mediated and planta transformation. Plant transformation involves three phases target gene, Plant tissues, vector for successful transformation. Plant transformation offers a momentous means to gain desire character or trait of interest. Plant transformation technique benefit agriculturalists to grow more crops in less area of land. And give more yield at less cost consumption. Plant transformation technology is familiarizing many crops with our desired characters. This review explains the natural method of plant transformation and benefits of transgenic plant.
Codon optimization is one of the key steps in achieving the high level expression of target gene. There are some key factors for consideration including transcription efficiency, translation efficiency, gene synthesis and protein folding.
DNA SEQUENCING METHODS AND STRATEGIES FOR GENOME SEQUENCINGPuneet Kulyana
This presentation will give you a brief idea about the various DNA sequencing methods and various strategies used for genome sequencing and much more vital information related to gene expression and analysis
Preparatiion of competent cells & Transformation Practical Sabahat Ali
Competence is ability of bacteria to take up foreign DNA.
Transformation is alteration of genetic material resulting from uptake of exogenous genetic material from surrounding environment through cell membrane .
Introduction
History
Cell culture techniques
Species cloned
Approaches of cell cloning
Monolayer culture- Dilution cloning
Microtitration plate
Suspension culture- Cloning in agar
Cloning in methocel
Isolation of clone
By clonal rings
By suspension clone
Application of cell cloning
Conclusion
Reference
Introduction.
Properties of Stem Cells.
Key Research events.
Embryonic Stem Cell.
Stem cell Cultivation.
Stem cells are central to three processes in an organism.
Research & Clinical Application of stem cell.
Research patents.
Conclusion.
Reference.
Plant transformation permits the introduction of the gene of interest for producing novel transgenic plants. "When a gene from one species is moved or relocated to another species by using recombinant DNA technology are called genetically modified organisms. Genetic engineering is one way to modify the plants by selecting for desired traits. Genetically modified organisms have foreign genes derive from not only plant source but also from bacteria, viruses, fungi, insects and animals. Transformation is the introduction and addition of the desired gene in plant for the generation of transgenic plant. Plant transformation is a challenging process for scientists. DNA transfer by artificial methods like DNA transfer through physical method is micro-injection, biolistic or gene gun methods, electroporation, silica carbide, microinjection, lipofection, microinjection. DNA also transfers by chemical methods. In natural method like in biological method, Agrobacterium-mediated transfer, Rhizobium, virus-mediated and planta transformation. Plant transformation involves three phases target gene, Plant tissues, vector for successful transformation. Plant transformation offers a momentous means to gain desire character or trait of interest. Plant transformation technique benefit agriculturalists to grow more crops in less area of land. And give more yield at less cost consumption. Plant transformation technology is familiarizing many crops with our desired characters. This review explains the natural method of plant transformation and benefits of transgenic plant.
Codon optimization is one of the key steps in achieving the high level expression of target gene. There are some key factors for consideration including transcription efficiency, translation efficiency, gene synthesis and protein folding.
DNA SEQUENCING METHODS AND STRATEGIES FOR GENOME SEQUENCINGPuneet Kulyana
This presentation will give you a brief idea about the various DNA sequencing methods and various strategies used for genome sequencing and much more vital information related to gene expression and analysis
Preparatiion of competent cells & Transformation Practical Sabahat Ali
Competence is ability of bacteria to take up foreign DNA.
Transformation is alteration of genetic material resulting from uptake of exogenous genetic material from surrounding environment through cell membrane .
Introduction
History
Cell culture techniques
Species cloned
Approaches of cell cloning
Monolayer culture- Dilution cloning
Microtitration plate
Suspension culture- Cloning in agar
Cloning in methocel
Isolation of clone
By clonal rings
By suspension clone
Application of cell cloning
Conclusion
Reference
Introduction.
Properties of Stem Cells.
Key Research events.
Embryonic Stem Cell.
Stem cell Cultivation.
Stem cells are central to three processes in an organism.
Research & Clinical Application of stem cell.
Research patents.
Conclusion.
Reference.
This is one of the major chapters for the examination NEET. A few questions are expected from this chapter and carry more weight as per the NEET syllabus.
genetic engineering: Genetic engineering, also called genetic modification, is the direct manipulation of an organism's genome using biotechnology. It is a set of technologies used to change the genetic makeup of cells, including the transfer of genes within and across species boundaries to produce improved or novel organisms. Many organism are manipulated with the help genetic engineering useful for mankind.
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.
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.
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.
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.
Model Attribute Check Company Auto PropertyCeline George
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This presentation provides a briefing on how to upload submissions and documents in Google Classroom. It was prepared as part of an orientation for new Sainik School in-service teacher trainees. As a training officer, my goal is to ensure that you are comfortable and proficient with this essential tool for managing assignments and fostering student engagement.
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It is possible to hide or invisible some fields in odoo. Commonly using “invisible” attribute in the field definition to invisible the fields. This slide will show how to make a field invisible in odoo 17.
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.
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptxEduSkills OECD
Andreas Schleicher presents at the OECD webinar ‘Digital devices in schools: detrimental distraction or secret to success?’ on 27 May 2024. The presentation was based on findings from PISA 2022 results and the webinar helped launch the PISA in Focus ‘Managing screen time: How to protect and equip students against distraction’ https://www.oecd-ilibrary.org/education/managing-screen-time_7c225af4-en and the OECD Education Policy Perspective ‘Students, digital devices and success’ can be found here - https://oe.cd/il/5yV
2. Recombinant DNA and
Gene Cloning
Recombinant DNA (rDNA) is a form of artificial
DNA that is created by combining two or more
DNA sequences originating from different
organism.
Recombinant DNA technology is a technology
which allows DNA to be produced via artificial
means. The procedure has been used to change
DNA in living organisms and may have even more
practical uses in the future.
3. Recombinant DNA technology is
one of the recent advances in
biotechnology, which was
developed by two scientists named
Boyer and Cohen in 1973.
4. Stanley N. Cohen (1935–) (top)
and Herbert Boyer (1936–)
(bottom), who constructed the
first recombinant DNA using
bacterial DNA and plasmids.
Stanley N. Cohen , who
received the Nobel Prize in
Medicine in 1986 for his
work on discoveries of
growth factors.
5. What is Recombinant DNA Technology?
Recombinant DNA technology is a
technology which allows DNA to be
produced via artificial means.
The procedure has been used to change
DNA in living organisms and may have even
more practical uses in the future.
It is an area of medical science that is just
beginning to be researched in a concerted
effort.
6. Recombinant DNA technology works by
taking DNA from two different sources and
combining that DNA into a single molecule.
That alone, however, will not do much.
Recombinant DNA technology only becomes
useful when that artificially-created DNA is
reproduced. This is known as DNA cloning.
9. Concept of Recombinant DNA
Recombinant DNA is a molecule that combines
DNA from two sources . Also known as gene
cloning.
Creates a new combination of genetic material
– Human gene for insulin was placed in bacteria
– The bacteria are recombinant organisms and
produce insulin in large quantities for diabetics
– Genetically engineered drug in 1986
Genetically modified organisms are possible
because of the universal nature of the genetic
code!
10. Genetic engineering is the application of
this technology to the manipulation of
genes. These advances were made
possible by methods for amplification of
any particular DNA segment( how? ),
regardless of source, within bacterial
host cells. Or, in the language of
recombinant DNA technology, the
cloning of virtually any DNA sequence
became feasible.
11. Six steps of Recombinant DNA
1. Isolating (vector and target gene)
2. Cutting (Cleavage)
3. Joining (Ligation)
4. Transforming
5. Cloning
6. Selecting (Screening)
13. Six basic steps are common to most
recombinant DNA experiments
1. Isolation and purification of DNA.
Both vector and target DNA molecules
can be prepared by a variety of
routine methods, which are not
discussed here. In some cases, the
target DNA is synthesized in vitro.
14. 2. Cleavage of DNA at particular sequences. As
we will see, cleaving DNA to generate
fragments of defined length, or with specific
endpoints, is crucial to recombinant DNA
technology. The DNA fragment of interest is
called insert DNA. In the laboratory, DNA is
usually cleaved by treating it with
commercially produced nucleases and
restriction endonucleases.
15. 3. Ligation of DNA fragments.
A recombinant DNA molecule is usually
formed by cleaving the DNA of interest to
yield insert DNA and then ligating the insert
DNA to vector DNA (recombinant DNA or
chimeric DNA). DNA fragments are
typically joined using DNA ligase (also
commercially produced).
– T4 DNA Ligase
16. 4. Introduction of recombinant DNA into
compatible host cells. In order to be
propagated, the recombinant DNA
molecule (insert DNA joined to vector
DNA) must be introduced into a
compatible host cell where it can replicate.
The direct uptake of foreign DNA by a host
cell is called genetic transformation (or
transformation). Recombinant DNA can
also be packaged into virus particles and
transferred to host cells by transfection.
17. 5. Replication and expression of
recombinant DNA in host cells.
Cloning vectors allow insert DNA to be
replicated and, in some cases, expressed
in a host cell. The ability to clone and
express DNA efficiently depends on the
choice of appropriate vectors and hosts.
18. 6. Identification of host cells that contain
recombinant DNA of interest. Vectors
usually contain easily scored genetic
markers, or genes, that allow the
selection of host cells that have taken up
foreign DNA. The identification of a
particular DNA fragment usually
involves an additional step—screening a
large number of recombinant DNA
clones. This is almost always the most
difficult step.
19. DNA cloning in a plasmid
vector permits amplification
of a DNA fragment.
21. 1. Analysis of Gene Structure and
Expression
2. Pharmaceutical Products
– Drugs
– Vaccines
3. Genetically modified organisms (GMO)
– Transgenic plants
– Transgenic animal
4. Application in medicine
22.
23. • Insulin
– Hormone required to
properly process sugars
and fats
– Treat diabetes
– Now easily produced by
bacteria
• Growth hormone
deficiency
– Faulty pituitary and
regulation
– Now easily produced by
bacteria
24. Genetically modified organisms (GMO)
Use of recombinant plasmids in
agriculture
– plants with genetically desirable
traits
• herbicide or pesticide resistant corn
& soybean
– Decreases chemical insecticide use
– Increases production
• “Golden rice” with beta-carotene
– Required to make vitamin A, which in
deficiency causes blindness
25. Crops have been
developed that are
better tasting, stay
fresh longer, and are
protected from disease
and insect infestations.
“Golden rice” has been
genetically modified to
contain beta-carotene
26.
27. Insect-resistant tomato plants
The plant on the left contains a gene that encodes a
bacterial protein that is toxic to certain insects that
feed on tomato plants. The plant on the right is a
wild-type plant. Only the plant on the left is able to
grow when exposed to the insects.
30. A transgenic
mouse
Mouse on right is
normal; mouse on
left is transgenic
animal expressing
rat growth hormone
31. Farm Animals and “Pharm”
Animals
These transgenic sheep
carry a gene for a
human blood protein
– This protein may help in
the treatment of cystic
fibrosis
32. Other benefits of GMOs
Disease resistance
There are many viruses, fungi, bacteria that cause plant
diseases
“Super-shrimp”
Cold tolerance
Antifreeze gene from cold water fish introduced to
tobacco and potato plants
Drought tolerance & Salinity tolerance
As populations expand, potential to grow crops in
otherwise inhospitable environments
33. What are restriction enzymes?
• Molecular scissors that cut double
stranded DNA molecules at specific
sequence that sequence is called as
recognition sequence or site .
• Found naturally in a wide variety of
prokaryotes
• An important tool for manipulating DNA.
• 3,000 enzymes have been identified,
around 200 have unique properties, many
are purified and available commercially
34. Discovery
• Arbor and Dussoix in 1962 discovered that
certain bacteria contain Endonucleases
which have the ability to cleave DNA.
• In 1970 Smith and colleagues purified and
characterized the cleavage site of a
Restriction Enzyme.
• Werner Arbor, Hamilton Smith and Daniel
Nathans shared the 1978 Nobel prize for
Medicine and Physiology for their
discovery of Restriction Enzymes.
35. Types of Restriction Endonucleases
Type I-Most complex and bifunctional i.e do restriction
as well as modification activity. Cleave DNA at random
site hence not used in rDNA technology. Eg Eco K,EcoB
TypeII-simple, cleave DNA at specific recognition
sequence site(Palindromic). There are more than 350
Type ii RE with 100 different recognition sites are uptill
known .They requires Mg 2+ Ions for cleavage. Only
Type II RE are used in rDNA technology.Eco R1,Hind III
Type III-These are intermediate between Type I & II
RE.Recognition site is asymmetrical sequence of 5-7
BP.Eg.EcoP1, EcoP15, Hint F3
36. Restriction Endonucleases nomenclature
Named for bacterial genus, species, strain, and type
Example: EcoR1
Genus: Escherichia
Species: coli
Strain: R
Order discovered: 1
Mechanism of Action
Restriction Endonuclease scan the length of the DNA , binds to the DNA
molecule when it recognizes a specific sequence and makes one cut in each
of the sugar phosphate backbones of the double helix – by hydrolyzing the
phoshphodiester bond. Specifically,the bond between the 3’ O atom and the P
atom is broken.
38. Restriction Endonucleases
Enzymes recognize specific 4-8 bp sequences
Some enzymes cut in a staggered fashion - “sticky ends”
Some enzymes cut in a direct fashion – “blunt ends”
39.
40.
41.
42.
43. In molecular cloning, a vector is a DNA molecule used as a
vehicle to artificially carry foreign genetic material into another
cell, where it can be replicated and/or expressed.
VECTOR
44.
45. CLONING VECTORS
•Cloning vectors are DNA molecules that are used to "transport" cloned
sequences between biological hosts and the test tube.
•Most vectors are genetically engineered.
•A vector is used to amplify a single molecule of DNA into many copes.
Cloning vectors share common properties:
1. Ability to replicate.
2. Easy to isolate and purify
3. Contain a genetic marker for selection.
4. Unique restriction sites to facilitate cloning of insert DNA.
5. Minimum amount of nonessential DNA to optimize cloning