Genetic engineering is the process of manually adding new DNA to an organism. The goal is to add one or more new traits that are not already found in that organism. Examples of genetically engineered (transgenic) organisms currently on the market include plants with resistance to some insects, plants that can tolerate herbicides, and crops with modified oil content
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.
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.
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.
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.
It is the basics of vector cloning which necessary for every and each student who is intrested in biotechnology. It is only starting, if you want to more than this then please comment on it.
this presentation gives information about cloning technique such as TOPO Cloning, SLIC and, Golden Gate Cloning.
این ارایه در مورد تکنیک های کلون کردن می باشد
Los días 7 y 8 de mayo organizamos en la Fundación Ramón Areces con la Fundación General CSIC el Simposio Internacional 'Microbiología: transmisión'. La "transmisión" en microbiología hace referencia al proceso por el que material genético es transferido de una célula a otra, de una población a otra. Es un proceso clave para entender el origen y la evolución de los seres vivos. El objetivo de esta reunión era conocer mejor la logística de la transmisión para ser capaces de modular o suprimir algunos procesos de transmisión dañinos.
It is the basics of vector cloning which necessary for every and each student who is intrested in biotechnology. It is only starting, if you want to more than this then please comment on it.
this presentation gives information about cloning technique such as TOPO Cloning, SLIC and, Golden Gate Cloning.
این ارایه در مورد تکنیک های کلون کردن می باشد
Los días 7 y 8 de mayo organizamos en la Fundación Ramón Areces con la Fundación General CSIC el Simposio Internacional 'Microbiología: transmisión'. La "transmisión" en microbiología hace referencia al proceso por el que material genético es transferido de una célula a otra, de una población a otra. Es un proceso clave para entender el origen y la evolución de los seres vivos. El objetivo de esta reunión era conocer mejor la logística de la transmisión para ser capaces de modular o suprimir algunos procesos de transmisión dañinos.
DNA cloning is a technique for reproducing DNA fragments.
It can be achieved by two different approaches:
▪ cell based
▪ using polymerase chain reaction (PCR).
a vector is required to carry the DNA fragment of interest into the host cell.
Now a day's these technique is tremendously use for in lab by using foreign Dna to to producing insulin in bacteria , plant with high yielding capacity by using Gene from another species
Study of cloning vectors and recombinant dna technologySteffi Thomas
Study of cloning vectors, restriction endonuclease and DNA ligase, Recombinant DNA technology, Application of genetic engineering in medicine, Application of rDNA technology and genetic engineering in the production of interferons, Vaccines-hepatitis-B, Hormones-Insulin, Brief introduction to PCR
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]
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
Biological screening of herbal drugs: Introduction and Need for
Phyto-Pharmacological Screening, New Strategies for evaluating
Natural Products, In vitro evaluation techniques for Antioxidants, Antimicrobial and Anticancer drugs. In vivo evaluation techniques
for Anti-inflammatory, Antiulcer, Anticancer, Wound healing, Antidiabetic, Hepatoprotective, Cardio protective, Diuretics and
Antifertility, Toxicity studies as per OECD guidelines
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
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The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
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Safalta Digital marketing institute in Noida, provide complete applications that encompass a huge range of virtual advertising and marketing additives, which includes search engine optimization, virtual communication advertising, pay-per-click on marketing, content material advertising, internet analytics, and greater. These university courses are designed for students who possess a comprehensive understanding of virtual marketing strategies and attributes.Safalta Digital Marketing Institute in Noida is a first choice for young individuals or students who are looking to start their careers in the field of digital advertising. The institute gives specialized courses designed and certification.
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Honest Reviews of Tim Han LMA Course Program.pptxtimhan337
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A Strategic Approach: GenAI in EducationPeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
Embracing GenAI - A Strategic ImperativePeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
4. DNA CLONING
– DNA cloning is a technique for reproducing DNA fragments.
– It can be achieved by two different approaches:
– ▪ cell based
– ▪ using polymerase chain reaction (PCR).
– a vector is required to carry the DNA fragment of interest into the host cell
5. – A single DNA molecule can be amplified allowing it to be:
Studied - Sequenced
Manipulated - Mutagenised or Engineered
Expressed - Generation of Protein
6. CLONING PROCESS
– Gene of interest is cut out with RE
– Host plasmid is cut with same RE
– Gene is inserted into plasmid and ligated with ligase
– New plasmid inserted into bacterium (transform)
7. PLASMID CLONING
STRATEGY
Involves five steps:
– Enzyme restriction digest of DNA sample.
– Enzyme restriction digest of DNA plasmid vector.
– Ligation of DNA sample products and plasmid vector.
– Transformation with the ligation products.
– Growth on agar plates with selection for antibiotic resistance.
11. STEP 4. TRANSFORMATION
OF LIGATION PRODUCTS
– The process of transferring exogenous DNA into cells is call “transformation”
– There are basically two general methods for transforming bacteria. The first is a
chemical method utilizing CaCl2 and heat shock to promote DNA entry into
cells.
– A second method is called electroporation based on a short pulse of electric
charge to facilitate DNA uptake.
15. TERMS USED IN CLONING
DNA recombination.
– The DNA fragment to be cloned is inserted into a vector.
Transformation.
– The recombinant DNA enters into the host cell and proliferates.
Selective amplification.
– A specific antibiotic is added to kill E. coli without any protection. The transformed E. coli is protected by the
antibiotic-resistance gene
16. CLONING VECTORS
Cloning vectors are DNA molecules that are used to "transport" cloned sequences
between biological hosts and the test tube.
– Cloning vectors share four common properties:
1. Ability to promote autonomous replication.
2. Contain a genetic marker (usually dominant) for selection.
3. Unique restriction sites to facilitate cloning of insert DNA.
4. Minimum amount of nonessential DNA to optimize cloning.
18. – Different types of cloning vectors are used for different types of cloning
experiments.
– The vector is chosen according to the size and type of DNA to be cloned.
19. PLASMID VECTORS
– Plasmid vectors are used to clone DNA ranging in size from several base pairs to
several thousands of base pairs (100bp -10kb).
– ColE1 based, pUC vehicles
– commercially available ones, eg pGEM3, pBlueScript
24. Why Plasmids are Good Cloning
Vectors
– small size (easy to manipulate and isolate)
– circular (more stable)
– replication independent of host cell
– several copies may be present (facilitates replication)
– frequently have antibiotic resistance (detection easy)
25. Disadvantages using plasmids
– Cannot accept large fragments
– Sizes range from 0.1- 10 kb
– Standard methods of transformation are inefficient
28. COSMID VECTOR
– Purpose:
Clone large inserts of DNA: size ~ 45 kb
– Features:
Cosmids are Plasmids with one or two Lambda
Cos sites.
– Presence of the Cos site permits in vitro packaging of cosmid DNA into Lambda
particles
29.
30. advantages of Lambda as Cloning
Vehicle:
– Strong selection for cloning of large inserts
– Infection process rather than transformation for entry of chimeric DNA into E.
coli host
31. Phagemid
A phagemid or phasmid is a plasmid that contains an f1 origin of replication from a
f1 phage.
It can be used as a type of cloning vector in combination with filamentous phage
M13.
A phagemid can be replicated as a plasmid, and also be packaged in viral particles.
35. RETROVIRAL VECTORS
– Retroviral vectors are used to introduce new or altered genes into the genomes of
human and animal cells.
– Retroviruses are RNA viruses.
– The viral RNA is converted into DNA by the viral reverse transcriptase and then is
efficiently integrated into the host genome
– Any foreign or mutated host gene introduced into the retroviral genome will be
integrated into the host chromosome and can reside there practically indefinitely.
– Retroviral vectors are widely used to study oncogenes and other human genes.
36.
37. EXPRESSION VECTORS
Allows a cloned segment of DNA to be translated into protein inside a bacterial or
eukaryotic cell.
– Vectors will contain the
– (a) in vivo promoter
– (b) Ampicillin selection
– (c) Sequencing primers
38.
39. advantages
– Produces large amounts of a specific protein
– Permits studies of the structure and function of proteins
– Can be useful when proteins are rare cellular components or difficult to isolate
40. REPORTER GENE VECTORS
– A gene that encodes a protein whose activity can be easily assayed in a cell in
which it is not normally expressed
– These genes are linked to regulatory sequences whose function is being tested
– Changes in transcriptional activity from the regulatory sequences are detected
by changes in the level of reporter gene expression
41. How is foreign DNA Inserted
into a Plasmid?
– To open up the DNA a restriction enzyme is used.
– Cut the DNA at a specific place called a restriction site.
– The result is a set of double-stranded DNA pieces with single-stranded ends
– These ends that jut out are not only "sticky" but they have gaps that can be now
be filled with a piece of foreign DNA
– For DNA from an outside source to bond with an original fragment, one more
enzyme is needed
– DNA ligase seals any breaks in the DNA molecule
42. RESTRICTION ENZYMES
A restriction enzyme (or restriction endonuclease) is an enzyme that cuts
double-stranded or single stranded DNA at specific recognition nucleotide
sequences known as restriction sites.
43.
44.
45.
46.
47. function:
– Inactivate foreign DNA
– Breaks only palindrome sequences, i.e. those exhibiting two-fold symmetry
– Important in DNA research, i.e. sequencing, hybridization
– Companies purify and market restriction enzymes
48. 5’ OVERHANGS
– 5' overhangs: The enzyme cuts asymmetrically within the recognition site such
that a short single-stranded segment extends from the 5' ends. Bam HI cuts in
this manner.
49. 3’ OVERHANGS
– 3' overhangs: Again, we see asymmetrical cutting within the recognition site,
but the result is a single-stranded overhang from the two 3' ends. KpnI cuts in
this manner.
50. BLUNT ENDS
– Blunts: Enzymes that cut at precisely opposite sites in the two strands of DNA
generate blunt ends without overhangs. SmaI is an example of an enzyme that
generates blunt ends.
51. Converting a 5’ overhang to
blunt end
– Both Klenow and T4 DNA polymerase can be used to fill in 5’ protruding ends
with dNTPs
– Used in joining DNA fragments with incompatible ends
– Once the ends have been blunted, ligation can proceed
52.
53. Converting a 3’ overhang to a
blunt end
– T4 DNA polymerase has a 3’-5’ exonuclease activity
– In the presence of excess dNTPs will convert a 3’ protruding end to a blunt end
– Ligation can know proceed