Recombinant DNA technology involves transferring genes between organisms using artificial means. It works by combining DNA from different sources into a single molecule. The process involves generating DNA fragments, inserting the fragments into vectors, introducing the vectors into host cells, and selecting clones containing the recombinant DNA. Common tools used include restriction enzymes to cut DNA, vectors like plasmids to carry DNA, bacterial hosts like E. coli, and techniques like transformation and selection to introduce and identify recombinant DNA. Applications include analyzing gene structure, producing pharmaceuticals, genetically modified organisms, and gene therapy.
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.
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.
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]
What are an expression vector? Detailed description of plant gene structure. Plant expression vector systems are generally consists of Ri and Ti plasmids.
The other vectors which are generally used are DNA and RNA viruses.
in this presentation, what are the steps and strategies involved the gene cloning and i was focused only on the 1st two steps of gene cloning.they are generation of foreign DNA molecules and selection of suitable vectors.
A DNA library is a collection of cloned restriction fragments of the DNA of an organism.
Two kinds of libraries will be discussed: genomic libraries and complementary DNA (cDNA) libraries.
Genomic libraries ideally contain a copy of every DNA nucleotide sequence in the genome.
In contrast, cDNA libraries contain those DNA sequences that appear as mRNA molecules, and these differ from one cell type to another.
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.
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]
What are an expression vector? Detailed description of plant gene structure. Plant expression vector systems are generally consists of Ri and Ti plasmids.
The other vectors which are generally used are DNA and RNA viruses.
in this presentation, what are the steps and strategies involved the gene cloning and i was focused only on the 1st two steps of gene cloning.they are generation of foreign DNA molecules and selection of suitable vectors.
A DNA library is a collection of cloned restriction fragments of the DNA of an organism.
Two kinds of libraries will be discussed: genomic libraries and complementary DNA (cDNA) libraries.
Genomic libraries ideally contain a copy of every DNA nucleotide sequence in the genome.
In contrast, cDNA libraries contain those DNA sequences that appear as mRNA molecules, and these differ from one cell type to another.
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.
This presentation deals with the introduction of Recombinant DNA Technology. The role of different enzymes. Specifically Restriction endonucleases and roles of various vectors.
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
A recombinant DNA molecule is produced by joining together two or more DNA segments usually originating from two different organisms.
More Specifically, a recombinant DNA molecule is a vector into which desired DNA fragment has been inserted to enable its cloning in an appropriate host.
Recombinant DNA molecules are produced with one of the following objectives:
1. To obtain large number of copies of specific DNA fragments.
2. Large scale production of the protein encoded by the gene.
3. Integration of the desired DNA fragment into target organism where it expresses itself.
Drought tolerant-genetically modified plants:
Present abiotic stress is a major challenge in our quest for sustainable food production as these may reduce the potential yields by 70% in crop plants
Of all abiotic stress, drought is regarded as the most damaging
Transgenic plants carrying genes for abiotic stress tolerance are being developed for water stress management
Conventional breeding approaches, involving inter specific and inter generic hybridizations and mutagenesis have been limited success.
Major problems have been the complexity of drought tolerance & low genetic yield components under drought conditions.
Unlike conventional plant breeding there is no need of repeated back crossing
Gene pyramiding or gene stacking through co-transformation of different genes with similar effects can be achieved.
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.
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
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Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
2. When a gene of one species is transferred to
another living organism by artificial means, it
is called recombinant DNA technology.
Also k/a genetic engineering.
Recombinant DNA technology works by
taking DNA from two different sources and
combining that DNA into a single molecule.
3. It occurs in following stages
Generation of DNA fragments & selection of
the desired piece of DNA (e.g. a human
gene).
Insertion of the selected DNA into a cloning
vector (e.g. a plasmid) to create a
recombinant DNA or chimeric DNA.
4. Introduction of the recombinant vectors
into
host cells (e.g. bacteria).
Multiplication & selection of clones
containing the recombinant molecules.
Expression of the gene to produce the
desired product.
5. Five Basic Steps in Gene Cloning
1. choose the appropriate DNA to be cloned,
genomic or cDNA.
2. Produce a collection of DNA fragments of
size suitable for inserting into appropriate
vectors.
3. Insert DNA fragments into the vector using
DNA ligase (DNA ligation.)
4. Introduce DNA fragments into a population
of bacteria (transformation.)
5. Select the colonies containing desired
sequence from the “library.”
11. Restriction endonucleases
DNA cutting enzymes:
Restriction endonucleases are one of the
most important groups of enzymes for the
manipulation of DNA.
These are the bacterial enzymes that can
cut/split DNA (from any source) at specific sites.
12. • They were first discovered in E.coli restricting
the replication of bacteriophages (virus), by
cutting the viral DNA (The host E. coli DNA is
protected from cleavage by addition of
methyl groups).
Thus, the enzymes that restrict the viral
replication are known as restriction enzymes
or restriction endonucleases.
13. Recognition sequence is the site where the DNA is
cut by a restriction endonuclease.
Restriction endonucleases can specifically
recognize DNA with a particular sequence of 4-8
nucleotides & cleave.
Cleavage patterns: Majority of restriction
endonucleases (particularly type II) cut DNA
at defined sites within recognition sequence.
14. The same sequence of bases is found on
both DNA strands, but in opposite order.
This arrangement is called a palindrome.
Palindromes are words or sentences that
read the same forward and backward.
15. The cut DNA fragments by restriction
endonucleases may have mostly sticky ends
(cohesive ends) or blunt ends.
DNA fragments with sticky ends are useful for
recombinant DNA experiments.
This is because the single-stranded sticky
DNA
ends can easily pair with any other DNA
fragment having complementary sticky ends.
16.
17.
18.
19. The cut DNA fragments are covalently
joined
together by DNA ligases.
These enzymes were originally isolated from
viruses.
They also occur in E.coli & eukaryotic cells.
DNA ligases actively participate in cellular
DNA repair process.
20. The hosts are the living systems or cells in
which the carrier of recombinant DNA
molecule or vector can be propagated.
There are different types of host cells-
prokaryotic (bacteria) & eukaryotic
(fungi, animals & plants).
21. Host cells, besides effectively
incorporating the vector's genetic
material, must be conveniently cultivated
in the laboratory to collect the products.
Microorganisms are preferred as host
cells, since they multiply faster compared
to cells of higher organisms (plants or
animals).
22. Escherichia coli:
Escherichia coli was the first organism used
in the DNA technology & continues to be
the host of choice by many workers.
The major drawback is that E. coli (or even
other prokaryotic organisms) cannot
perform post-translational modifications.
Bacillus subtilis as an alternative to E.coli.
23. The most commonly used eukaryotic organism
is the yeast, Saccharomyces cerevisiae.
Certain complex proteins which cannot be
synthesized by bacteria can be produced by
mammalian cells e.g. tissue plasminogen activator.
The mammalian cells possess the machinery to
modify the protein to the active form (post-
translational modifications).
24. Vectors are the DNA molecules, which
can carry a foreign DNA fragment to be
cloned.
They are self-replicating in an appropriate
host cell.
The most important vectors are plasmids,
bacteriophages, cosmids & artificial
chromosome vectors.
25. Ideal vector
A vector should have four characteristics:
1.Ability to replicate independently of the host cell
2.A recognition sequence for a restriction enzyme
(cloning site)
3.One or more selectable/reporter genes
4.Small size in comparison with host’s
chromosomes
26. Plasmids are extrachromosomal,
double- stranded, circular, self-
replicating DNA molecules.
Almost all bacteria have plasmids.
Size of plasmids varies from 1 to 500 kb.
Plasmids contribute to about 0.5 to 5.0%
of total DNA of bacteria.
27. pBR322 has a DNA sequence of 4,361 bp.
It carries genes resistance for ampicillin (Amp1) &
tetracycline (Tel1) that serve as markers for the
identification of clones carrying plasmids.
The plasmid has unique recognition sites for the
action of restriction endonucleases - EcoRl,
Hindlll, BamHl, Sall & Pstll
28.
29. The other plasmids employed as cloning
vectors include pUC19 (2,686 bp, with
ampicillin resistance gene) & derivatives
of pBR322-pBR325, pBR328 & pBR329.
30. Bacteriophages or phages are the viruses
that replicate within the bacteria.
In case of certain phages, their DNA gets
incorporated into the bacterial chromosome
& remains there permanently.
31. Phages can take up larger DNA segments
than plasmids.
Phage vectors are preferred for working
with genomes of human cells.
The most commonly used phages are
bacteriophage λ (phage λ) &
bacteriophage (phage M13).
32. Cosmids are vectors possessing the characteristics of both
plasmid & phage λ.
Cosmids can be constructed by adding a fragment of
phage λ DNA including Cos site, to plasmids.
A foreign DNA (about 40 kb) can be inserted into cosmid
DNA .
The recombinant DNA, formed can be packed as phages &
injected into E.coli.
Inside host cell, cosmids behave like plasmids & replicate &
can carry larger fragments of foreign DNA
33. Human artificial chromosome (HAC):
Artificial chromosome is a synthetically
produced vector DNA, possessing the
characteristics of human chromosome.
HAC may be considered as a self-replicating
microchromosome with a size ranging from 1/10th
to 1/5th of a human chromosome.
It can carry long human genes.
34. Yeast artificial chromosome (YAC) is a
synthetic DNA that can accept large
fragments of foreign DNA (particularly
human DNA).
It is possible to clone large DNA pieces
by
using YAC.
35. Construction of BACs is based on one F-
plasmid which is larger than the other
plasmids used as cloning vectors.
BACs can accept DNA inserts of around 300
kb.
36. Transformation:
Transformation is the method of
introducing
foreign DNA into bacterial cells (e.g. E.coli).
Uptake of plasmid DNA by E.coli is carried
out in ice-cold CaCl2 (0-5˚C) & a
subsequent
heat shock (37-45˚C for about 90 sec).
37. A natural microbial recombination process.
During conjugation, two live bacteria (a
donor & a recipient) come together, join by
cytoplasmic bridges & transfer single
stranded DNA (from donor to recipient).
In side recipient cell, new DNA may
integrate
with the chromosome or may remain free.
38. It is a technique involving electric field
mediated membrane permeabiIization.
Electric shocks can also induce cellular
uptake of exogenous DNA (believed to be
via the pores formed by electric pulses) from
the suspending solution.
It is a simple & rapid technique for
introducing genes into cells.
39. Liposomes are circular lipid molecules,
which have an aqueous interior that can
carry nucleic acids.
Several techniques have been developed
to encapsulate DNA in liposomes.
The liposome mediated gene transfer is
referred to as lipofection.
40. Treatment of DNA fragment with liposomes,
DNA pieces get encapsulated inside liposomes.
These liposomes can adhere to cell
membranes
& fuse with them to transfer DNA fragments.
The DNA enters the cell & to the nucleus.
Positively charged liposomes efficiently
complex with DNA, bind to cells & transfer DNA
41. It is possible to directly transfer the DNA into
the cell nucleus.
Microinjection & particle bombardment
are the two techniques used for this
purpose.
42. Selection and screening Identification of host cells containing
recombinant DNA requires genetic selection
or screening or both.
In a selection, cells are grown under
conditions in which only transformed cells
can survive; all the other cells die.
In contrast, in a screen, transformed cells
have to be individually tested for the
presence of the desired recombinant DNA.
Many selection strategies involve selectable
marker genes— genes whose presence can
easily be detected or demonstrated
43.
44. Screening (Strategies)1. Gel Electrophoresis Allows Separation of
Vector DNA from Cloned Fragments
2. Cloned DNA Molecules Are Sequenced
Rapidly by the Dideoxy Chain-Termination
Method
3. The Polymerase Chain Reaction Amplifies
a Specific DNA Sequence from a Complex
Mixture
4. Blotting Techniques Permit Detection of
Specific DNA Fragments and mRNAs with
DNA Probes
45.
46. 1. Analysis of Gene Structure and Expression
2. Pharmaceutical Products
Drugs
Vaccines
1. Genetically modified organisms (GMO)
Transgenic plants
Transgenic animal
1. Application in medicine
- Gene therapy
47. Analysis of Gene Structure and
Expression
Using specialized recombinant DNA
techniques, researchers have determined
vast amounts of DNA sequence including the
entire genomic sequence of humans and
many key experimental organisms.
This enormous volume of data, which is
growing at a rapid pace, has been stored
and organized in the GenBank at the National
Institutes of Health, Bethesda, Maryland
48. Pharmaceutical Products Some pharmaceutical applications of DNA
technology:
Large-scale production of human hormones
and other proteins with therapeutic uses
Production of safer vaccines
A number of therapeutic gene products —
insulin, the interleukins, interferons, growth
hormones, erythropoietin, and coagulation
factor VIII—are now produced commercially
from cloned genes
49.
50. 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
51. 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
52. 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.