Recombinant DNA technology uses restriction enzymes and DNA ligase to cut and paste genes between organisms. It has led to important applications like producing human insulin from bacteria, creating genetically modified crops with desirable traits, and developing new vaccines and pharmaceuticals. The basic steps involve isolating a gene, inserting it into a vector, transforming host cells, and identifying recombinant cells that express the gene of interest. This technique has generated unlimited copies of genes and advanced fields like gene therapy, forensics, and biofuel production.
Visual Merchandising is the activity of maximizing the impact of Retail Display space and has become an important element in retailing. Visual Merchandisers create displays using color, lighting, space, product information, sensory inputs such as smell, touch, and sound as well as technologies such as digital displays and interactive installations. The objective of this highly practical program is to ensure that attendees are furnished with the core concepts and skills used in visual merchandising. Hamstech course will enable students to identify, select and pursue their career opportunities in the retail industry. The retail sector is a major employer and requires skilled staff in a range of jobs such as: Shop Floor Planning and Display, Window Display, Sales, Buying, Staff Management and Supervision.
The Definition of Marketing Series:Relationship Marketing. Learn how to use relationship marketing to improve your firm's performance. Relationship marketing is also a key factor underpinning success in social media marketing as it builds engagement with customers. For more information go to: http://hausmanmarketresearch.org
In this Slideshare i had define about product and services.
difference between product and services.
types of product and services.
if you like my presentation do like and follow.
Thank you
Visual Merchandising is the activity of maximizing the impact of Retail Display space and has become an important element in retailing. Visual Merchandisers create displays using color, lighting, space, product information, sensory inputs such as smell, touch, and sound as well as technologies such as digital displays and interactive installations. The objective of this highly practical program is to ensure that attendees are furnished with the core concepts and skills used in visual merchandising. Hamstech course will enable students to identify, select and pursue their career opportunities in the retail industry. The retail sector is a major employer and requires skilled staff in a range of jobs such as: Shop Floor Planning and Display, Window Display, Sales, Buying, Staff Management and Supervision.
The Definition of Marketing Series:Relationship Marketing. Learn how to use relationship marketing to improve your firm's performance. Relationship marketing is also a key factor underpinning success in social media marketing as it builds engagement with customers. For more information go to: http://hausmanmarketresearch.org
In this Slideshare i had define about product and services.
difference between product and services.
types of product and services.
if you like my presentation do like and follow.
Thank you
This presentation gives a brief information about the Seven P's.
The four P's are also know as the traditional P's and the other three P's are known as the modern or extended P's.
3.1 Key control and key handling procedures
3.2 Mail and message handling
3.3 Paging and luggage handling
3.4 Rules of the house (for guest and staff) & Black list
3.5 Bell Desk and Concierge
Technologies: Use of Technologies in retailing - Electronic Data Interchange (EDI), Radio Frequency Identification (RFI), Data Base Management system, E-Retailing: Formats, Challenges, Green Retailing Concept, Importance of Green Retailing.
This presentation gives a brief information about the Seven P's.
The four P's are also know as the traditional P's and the other three P's are known as the modern or extended P's.
3.1 Key control and key handling procedures
3.2 Mail and message handling
3.3 Paging and luggage handling
3.4 Rules of the house (for guest and staff) & Black list
3.5 Bell Desk and Concierge
Technologies: Use of Technologies in retailing - Electronic Data Interchange (EDI), Radio Frequency Identification (RFI), Data Base Management system, E-Retailing: Formats, Challenges, Green Retailing Concept, Importance of Green Retailing.
The ability to recreate computational results with minimal effort and actionable metrics provides a solid foundation for scientific research and software development. When people can replicate an analysis at the touch of a button using open-source software, open data, and methods to assess and compare proposals, it significantly eases verification of results, engagement with a diverse range of contributors, and progress. However, we have yet to fully achieve this; there are still many sociotechnical frictions.
Inspired by David Donoho's vision, this talk aims to revisit the three crucial pillars of frictionless reproducibility (data sharing, code sharing, and competitive challenges) with the perspective of deep software variability.
Our observation is that multiple layers — hardware, operating systems, third-party libraries, software versions, input data, compile-time options, and parameters — are subject to variability that exacerbates frictions but is also essential for achieving robust, generalizable results and fostering innovation. I will first review the literature, providing evidence of how the complex variability interactions across these layers affect qualitative and quantitative software properties, thereby complicating the reproduction and replication of scientific studies in various fields.
I will then present some software engineering and AI techniques that can support the strategic exploration of variability spaces. These include the use of abstractions and models (e.g., feature models), sampling strategies (e.g., uniform, random), cost-effective measurements (e.g., incremental build of software configurations), and dimensionality reduction methods (e.g., transfer learning, feature selection, software debloating).
I will finally argue that deep variability is both the problem and solution of frictionless reproducibility, calling the software science community to develop new methods and tools to manage variability and foster reproducibility in software systems.
Exposé invité Journées Nationales du GDR GPL 2024
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Phenomics assisted breeding in crop improvementIshaGoswami9
As the population is increasing and will reach about 9 billion upto 2050. Also due to climate change, it is difficult to meet the food requirement of such a large population. Facing the challenges presented by resource shortages, climate
change, and increasing global population, crop yield and quality need to be improved in a sustainable way over the coming decades. Genetic improvement by breeding is the best way to increase crop productivity. With the rapid progression of functional
genomics, an increasing number of crop genomes have been sequenced and dozens of genes influencing key agronomic traits have been identified. However, current genome sequence information has not been adequately exploited for understanding
the complex characteristics of multiple gene, owing to a lack of crop phenotypic data. Efficient, automatic, and accurate technologies and platforms that can capture phenotypic data that can
be linked to genomics information for crop improvement at all growth stages have become as important as genotyping. Thus,
high-throughput phenotyping has become the major bottleneck restricting crop breeding. Plant phenomics has been defined as the high-throughput, accurate acquisition and analysis of multi-dimensional phenotypes
during crop growing stages at the organism level, including the cell, tissue, organ, individual plant, plot, and field levels. With the rapid development of novel sensors, imaging technology,
and analysis methods, numerous infrastructure platforms have been developed for phenotyping.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
Salas, V. (2024) "John of St. Thomas (Poinsot) on the Science of Sacred Theol...Studia Poinsotiana
I Introduction
II Subalternation and Theology
III Theology and Dogmatic Declarations
IV The Mixed Principles of Theology
V Virtual Revelation: The Unity of Theology
VI Theology as a Natural Science
VII Theology’s Certitude
VIII Conclusion
Notes
Bibliography
All the contents are fully attributable to the author, Doctor Victor Salas. Should you wish to get this text republished, get in touch with the author or the editorial committee of the Studia Poinsotiana. Insofar as possible, we will be happy to broker your contact.
DERIVATION OF MODIFIED BERNOULLI EQUATION WITH VISCOUS EFFECTS AND TERMINAL V...Wasswaderrick3
In this book, we use conservation of energy techniques on a fluid element to derive the Modified Bernoulli equation of flow with viscous or friction effects. We derive the general equation of flow/ velocity and then from this we derive the Pouiselle flow equation, the transition flow equation and the turbulent flow equation. In the situations where there are no viscous effects , the equation reduces to the Bernoulli equation. From experimental results, we are able to include other terms in the Bernoulli equation. We also look at cases where pressure gradients exist. We use the Modified Bernoulli equation to derive equations of flow rate for pipes of different cross sectional areas connected together. We also extend our techniques of energy conservation to a sphere falling in a viscous medium under the effect of gravity. We demonstrate Stokes equation of terminal velocity and turbulent flow equation. We look at a way of calculating the time taken for a body to fall in a viscous medium. We also look at the general equation of terminal velocity.
1. BASIC PRINCIPLE OF RECOMBINANT DNA
TECHNOLOGY AND ITS APPLICATIONS
Presented by:
Prashanth M E
1ST Sem, M Pharm
Department of Pharmacology
Government College of Pharmacy,
Bengaluru
Guided by:
Dr. Rajesh M S
Assistant Professor
Department of Pharmacology
Government College of Pharmacy, Bengaluru
2. MOLECULAR BIOTECHNOLOGY
It is an scientific technique used to transfer specific units of genetic information from
one organism to another.
The field of modern biotechnology started when recombinant human insulin
produced by bacteria was first marketed in the United States in 1982.
The effort leading to this landmark began in the early 1970s when research
scientists developed protocols to construct new types of bacterial plasmids or
vectors, by cutting out and pasting pieces of DNA together to create a new piece
of DNA (recombinant DNA) that could be inserted into host bacterium such as
E.coli.
3. RECOMBINANT DNA TECHNOLOGY
It is the art of cutting and pasting genes.
Paul Berg, Herbert Boyer, Annie Change, and Stanley Cohen are the scientists made
the first recombinant DNA molecule in 1973.
This technique uses a number of tools that to construct new combinations of
DNA (recombinant DNA or rDNA) in the laboratory for different purposes.
The rDNA molecule thus constructed can be introduced into an appropriate host
cell, where it can be multiplied and generate many copies. This forms the basic
concept of the process known as gene cloning or DNA cloning
Gene cloning can generate unlimited copies of a DNA molecule (e.g.,
recombinant DNA) by replication in a host cell.
4. The DNA from a donor organism is extracted,
enzymatically and joined (ligated) to another DNA
entity to form a new, recombined DNA molecule.
This cloning vector–insert DNA construct is
transferred and maintained within a host cell. It is
called transformation.
•Those host cells that take up the DNA construct are
identified and selected from those that do not.
•If required, a DNA construct can be created so that
the protein product encoded by the cloned DNA
sequence is produced in the host cell.
5. TOOLS OF RECOMBINANT DNA TECHNOLOGY
Cloned DNA and a vehicle DNA to carry the DNA into a host cell and multiply.
The process of making a recombinant DNA requires the precise cutting and stitching
of DNA molecules,
This involves a number of molecular tools—the enzymes to cut and modify the DNA,
finally resulting in a recombinant DNA molecule.
The major enzymes required for the making of an rDNA molecule are:
Type II restriction endonucleases
DNA ligase
Alkaline phosphatase.
6. 1. RESTRICTION ENDONUCLEASES
First isolated in 1968 by a group of three scientists—H.O. Smith, K.W. Wilcox, and
T.J. Kelley
These enzymes are also known as molecular scissors.
One of the first type II restriction endonucleases to be characterized was from the
bacterium Escherichia coli, and it was originally designated EcoRI.
It is a homodimeric protein that binds to a DNA region with a specific palindromic
sequence. The EcoRI recognition sequence consists of 6 base pairs (bp) and is cut
between the guanine and adenine residues on each strand.
7. Restriction Enzyme Nomenclature
The very name of the restriction enzymes consists of three parts:
• An abbreviation of the genus and the species of the organism to 3 letters, for
example- Eco for Escherichia coli identified by the first letter, E, of the genus and
the first two letters, co, of the species.
• It is followed by a letter, number or combination of both of them to signify the
strain of the species.
• A Roman numeral to indicate the order in which the different restriction-
modification systems were found in the same organism or strain
8.
9. This enzyme is responsible for the formation of the phosphodiester linkage
between two adjacent nucleotides and thus joins two double-stranded DNA
fragments.
2. DNA LIGASE
10. 3. ALKALINE PHOSPHATASE
Self-ligation is the process of annealing the sticky ends
of the linearized vector without inserting the foreign
DNA fragment.
This can be prevented by using alkaline phosphatase.
It removes the phosphate group from the 5′ end of the
DNA molecule resulting in a 5′OH group.
bacterial alkaline phosphatase—BAP
calf intestine alkaline phosphatase—CAP
11.
12. VECTORS: THE VEHICLE FOR CLONING
Vectors act as a vehicle for carrying foreign DNA into a host cell for multiplication.
ORIGIN OF REPLICATION
SELECTABLE MARKERS
o positive selection - antibiotic resistance genes
o negative selection - insertional inactivation.
MULTIPLE CLONING SITES (MCS) OR POLYLINKER
SMALL SIZE
15. b) BACTERIOPHAGE-CLONING VECTORS
The viruses that infect bacteria are called bacteriophages. viruses usually contain a
comparatively small DNA genome surrounded by a protein coat.
Phage-cloning Vectors
DNA
replication
Packagin
g
Assembly
Infect E. coli
16. M13-based Cloning Vectors
Derived from bacteriophage M13.
It is lysogenic filamentous phage with a circular
single-stranded DNA genome about 6,407 bp
(6.4 kb) in length
The advantages of M13-based vectors are that they contain the same polylinker and
alpha-peptide fragments as the pUC series and recombinants can be selected by the
blue → white color test.
17. c) COSMID-CLONING VECTORS
A cosmid can be defined as a plasmid that contains
a cos site from the lambda phage genome.
The simplest cosmid vector has a ColE1 origin of
replication, selectable markers (Ampr), and
suitable polylinker sites and lambda cos site.
Ligation of the cosmid vector and foreign DNA
fragments of sizes up to 45 kb is similar to
ligation into a lambda substitution vector.
18. d) YEAST ARTIFICIAL CHROMOSOMES (YACS)
The vectors that enable the formation of
artificial chromosomes with the foreign DNA
fragments and cloning into yeast. These are
used for the cloning of very large DNA
fragments in the range of 500 to 1,000 kb.
19. e) BACTERIAL ARTIFICIAL CHROMOSOMES (BACS)
These are the cloning vectors based on the extrachromosomal plasmids of E. coli
called F factor or fertility factor. These vectors enable the construction of artificial
chromosomes, which can be cloned in E. coli.
This vector is useful for cloning DNA fragments up to 350 kb, but can be handled
like regular bacterial plasmid vectors, and is very useful for sequencing large
stretches of chromosomal DNA. Like any other vector, BACs contain ori
sequences derived from E. coli plasmid F factor, multiple cloning sites (MCS)
having unique restriction sites, and suitable selectable markers.
20. f) ANIMAL AND PLANT VECTORS (SHUTTLE VECTORS)
There are some vectors developed for transforming plant and animal cells.
These are often called shuttle vectors as they replicate in both prokaryotic and
eukaryotic hosts.
The common features of such shuttle vectors
o Capable of replicating in two or more types of hosts including prokaryotic and
eukaryotic cells.
o Replicate autonomously, or integrate into the host genome and replicate when the
host cell multiplies.
o Commonly used for transporting genes from one organism to another (i.e.,
transforming animal and plant cells).
21. MAMMALIAN VECTORS
These are shuttle vectors developed for use in mammalian tissue culture.
These eukaryotic origins of replication are typically derived from well-
characterized mammalian viruses such as simian virus 40 (SV-40) with sv-40 ori
and large tantigen system or Epstein-Barr virus
In addition to the origin of replication, these shuttle vectors also carry antibiotic
resistance genes, which function in eukaryotic cells
22. PLANT VECTORS
These shuttle vectors are based on certain viruses and bacteria, which are
pathogenic to plants. Plant viruses such as the cauliflower mosaic virus (CMV),
tobacco mosaic virus (TMV), and Gemini viruses were used for developing vectors
for plant cell transformations, but with limited success.
The most successful shuttle vectors developed for the plant system are those that
are based on the Ti plasmid of agrobacterium tumifaciens, a bacterium that causes
tumor formation in plants.
23. HOST CELLS
The host cell can be bacteria, yeast, plant or animal cells.
E. coli is the most widely used organism in rDNA experiments.
• It is very simple, easy to handle, grows rapidly and is able to accept and maintain a
range of vectors.
• The doubling time of e. coli under ideal growth conditions is 20 minutes.
• As the cell undergoes multiplication, the rDNA within the cell also undergoes
multiplication independent of its genome.
If the recombinant protein that we want to produce is of eukaryotic origin, it is better
to opt for a eukaryotic host system such as yeast.
• Yeasts are the simplest of the eukaryotic systems, single-celled, genetically and
physiologically well-characterized, and easy to grow and manipulate.
24. MAKING RECOMBINANT DNA
• The isolation and purification of vectors
and the DNA fragments containing the
gene to be cloned.
o Digest the vector DNA with a suitable
restriction endonuclease and make it
linear with or without sticky ends
o Isolate the DNA fragment carrying the
gene by digesting the genome
o The sticky ends of the two DNA strands
come closer by the base
complementation and become hydrogen
bonded to each other.
25. TRANSGENICS—INTRODUCTION OF RECOMBINANT
DNA INTO HOST CELLS
Transformation
• Cells take up foreign
DNA from their
environment.
• Chemical treatments
can enhance the ability
of cells to take up the
foreign DNA
Transfection
• rDNA is mixed with
charged substances
such as calcium
phosphate, cationic
liposomes, and
overlayered on the
host cells.
• Uptake of the external
DNA by these host
cells.
Electroporation
• Electric current is used
to create transient
microscopic pores in
the cell membrane of
the host cell.
• Temporary openings
foreign DNA enters the
cell.
Microinjection
• DNA fragment or is
directly injected into
the nucleus of plant
and animal cells.
• Carried out without the
use of any specialized
vector.
• Help of a glass
microinjection tube or
syringe.
The biolistic method
• Introducing foreign
DNA into plant cells
with the help of a gene
gun.
• Microscopic particles
of gold or tungsten
coated with the DNA
of interest is
bombarded into the
cells at a high velocity.
26. IDENTIFICATION OF RECOMBINANTS
During the process of genetic transformation only a very low percentage of the total
cell population takes or receives the recombinant DNA.
In most of the cases there are two stages of selection.
First, one is the selection of transformed cells
The second one is to identify the transformed cells that have the recombinant
plasmid
The transformed cells contain a plasmid and can be identified by the
• Positive selection method - antibiotic-resistance genes
• Negative selection method – insertional activation
27. STEPS IN RECOMBINANT DNA TECHNOLOGY
(i) Selection and isolation of DNA insert
(ii)Selection of suitable cloning vector
(iii) Introduction of DNA-insert into vector
to form rDNA molecule
(iv) rDNA molecule is introduced into a
suitable host
(v) Selection of transformed host cells
(vi) Expression and Multiplication of DNA
insert in the host
28. APPLICATIONS
Production of Transgenic Plants:
For qualities like resistance to herbicides, insects or viruses or with expression of male sterility
etc.
Production of Transgenic Animals
• To increase the speed and range of selective breeding in case of animals
• For the production of better farm animals so as to ensure more commercial benefits.
• Production of certain proteins and pharmaceutical compounds.
• To study the gene functions in different animal species.
29. Production of Hormones:
• Bacterial cells like E.coli are utilized for the production of different fine chemicals like
insulin, somatostatin, somatotropin and p-endorphin.
• Human Insulin Hormone i.e., Humulin is the first therapeutic product which was produced
by the application of rec DNA technology.
Biosynthesis of Interferon:
• The gene of human fibroblasts is inserted into the bacterial plasmid.
• These genetically engineered bacteria are cloned and cultured so that the gene is expressed
and the interferons are produced in fairly high quantities.
• This interferon, so produced, is then extracted and purified.
30. Production of Vaccines:
• A number of vaccines have been synthesized through rDNA technology, which are effective
against numerous serious diseases caused by bacteria, viruses or protozoa.
• These include vaccines for polio, malaria, cholera, hepatitis, rabies, smallpox, etc.
• DNA-vaccine is the preparation that contains a gene encoding an immunogenic protein from
the concerned pathogen.
Production of Antibiotics:
• rDNA technology helps in increasing the production of antibiotics by improving the
microbial strains through modification of genetic characteristics.
• Some important antibiotics are tetracycline, penicillin, streptomycin, novobiocin,
bacitracin, etc.
31. Production of Commercially Important Chemicals:
• alcohols and alcoholic beverages obtained through fermentation
• organic acids like citric acid, acetic acid, etc. and vitamins produced by microorganisms.
Application in Enzyme Engineering:
• the enzymes are encoded by genes, and if there are changes in a gene then the enzyme
structure also changes.
• Enzyme engineering utilizes the same fact and can be explained as the modification of an
enzyme structure by inducing alterations in the genes which encode for that particular
enzyme.
32. Prevention and Diagnosis of Diseases:
• Monoclonal antibodies are useful tools for disease diagnosis.
• Monoclonal antibodies are produced by using the technique called hybridoma technology
• rDNA provides methods for the. prevention of a number of diseases like AIDS, cholera, etc.
Gene Therapy:
• Gene therapy is undoubtedly the most beneficial area of genetic engineering for human
beings.
• It involves delivery of specific genes into human body to correct the diseases. Thus, it is the
treatment of diseases by transfer and expression of a gene into the patients’ cells so as to
ensure the restoration of a normal cellular activity.
33. Applications in forensic science:
• The applications of rec DNA technology in forensic sciences depends on the technique
called DNA profiling or DNA fingerprinting.
• It enables us to identify any person by analysing his hair roots Wood stains, serum, etc.
• DNA fingerprinting also helps to solve the problems of parentage and to identify the
criminals.
Biofuel Production:
• Biofuels are derived from biomass and these are renewable and cost effective.
• Genetic engineering plays an essentially important role in a beneficial and largescale
production of biofuels like biogas. bio hydrogen biodiesel bio-ethanol., etc.
• Genetic engineering helps to improve organisms for obtaining higher product yields and
product tolerance.
34. Practical Applications of Genetic Engineering:
rDNA technology has an immense scope in Research and Experimental studies.
It is applied for:
a. Localizing specific genes.
b. Sequencing of DNA or genes.
c. Study of mechanism of gene regulation.
d. Molecular analysis of various diseases.
e. Study of mutations in DNA, etc.
35. REFERENCE
1. Glick BR, Pasternak JJ, Patten CL. Molecular biotechnology: Principles and
applications of recombinant DNA. 4th ed. Pasternak JJ, Glick BR, Patten CL, editors.
Washington, D.C., DC: American Society for Microbiology; 2009.
2. Nair AJ. Introduction to biotechnology and genetic engineering. New Delhi, India:
Laxmi Publications; 2008.
3. Satyanarayana U. Biochemistry - E-Book. 5th ed. New Delhi, India: Elsevier; 2017
4. Nagel KM. Principles of recombinant DNA technology. In: Introduction to Biologic
and Biosimilar Product Development and Analysis. Cham: Springer International
Publishing; 2018. p. 1–29.
5. S.A. shinde et.al, recombinant DNA technology and its applications: A review,
international journal of medipharm research p.79-88