BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Bacteriophage vectors
Bacteriophage
WHY BACTERIOPHAGE AS A VECTOR?
M13 phage
Genome of m13 phage
Life cycle and dna replication of m13
CONSTRUCTION M13 AS PHAGE VECTOR
M13 MP 2 vector
M13MP7 VECTOR
Selection of recombinants
Lambda replacement vectors
LAMBDA EMBL 4 VECTOR
P1 PHAGE
GENOME OF P1 PHAGE
P1 PHAGE AS VECTOR
P1 phage vector system
This presentation covers a general introduction to expression vector, its components, types, and its application. Then it covers some of the expression system with examples.
molecular biology phage vector, full lifecycle and all necessary information regarding lambda phage, it contain 2 types that is insertion and replacement.
Bacteriophage vectors
Bacteriophage
WHY BACTERIOPHAGE AS A VECTOR?
M13 phage
Genome of m13 phage
Life cycle and dna replication of m13
CONSTRUCTION M13 AS PHAGE VECTOR
M13 MP 2 vector
M13MP7 VECTOR
Selection of recombinants
Lambda replacement vectors
LAMBDA EMBL 4 VECTOR
P1 PHAGE
GENOME OF P1 PHAGE
P1 PHAGE AS VECTOR
P1 phage vector system
This presentation covers a general introduction to expression vector, its components, types, and its application. Then it covers some of the expression system with examples.
molecular biology phage vector, full lifecycle and all necessary information regarding lambda phage, it contain 2 types that is insertion and replacement.
Genetic transformation & success of DNA ligation Sabahat Ali
DNA is ligated through DNA Ligase, problems may occur during DNA ligation are
1) vector cyclization
2) vector-vector concatemers
3) target DNA-target DNA ligation
Transgenic animal production and its applicationkishoreGupta17
A genetically modified animal with the heterologous gene of interest being inserted for the purpose of biopharming or make a diseased model to study the consequences of disease and its probable therapy
for cloning and expression of exogenous gene or gene throthrough vector it must be introduced into the host cell through transformation , ,transduction, electroporation gene gun etc.
A genetically engineered DNA molecule from bacteria , phage or yeast to carry foreign DNA for the purpose of cloning and expression of the inserted DNA of interest in RDT
A cytological technique to detect the nature of adjacent chromosomal regions by using different staining technique assisted with some pre treatment of metaphase chromosomes prepared on the slides
The Indian economy is classified into different sectors to simplify the analysis and understanding of economic activities. For Class 10, it's essential to grasp the sectors of the Indian economy, understand their characteristics, and recognize their importance. This guide will provide detailed notes on the Sectors of the Indian Economy Class 10, using specific long-tail keywords to enhance comprehension.
For more information, visit-www.vavaclasses.com
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
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.
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.
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.
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.
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.
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
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
This is a presentation by Dada Robert in a Your Skill Boost masterclass organised by the Excellence Foundation for South Sudan (EFSS) on Saturday, the 25th and Sunday, the 26th of May 2024.
He discussed the concept of quality improvement, emphasizing its applicability to various aspects of life, including personal, project, and program improvements. He defined quality as doing the right thing at the right time in the right way to achieve the best possible results and discussed the concept of the "gap" between what we know and what we do, and how this gap represents the areas we need to improve. He explained the scientific approach to quality improvement, which involves systematic performance analysis, testing and learning, and implementing change ideas. He also highlighted the importance of client focus and a team approach to quality improvement.
2. • Plasmid has carrying capacity of maximum 10
kb insert
• For DNA library DNA size needed is large
• So virus is used as vector
• viral vectors are genetically engineered to
carry the cloned genomic DNA /c DNA
• The basis of gene introduction into the host
cells lies in the process of Transduction via
which a virus infect and inject the viral DNA
3. Natural Lambda Phage
• λPhage is a temperate phage that infects E.coli and
replicates by lytic & lysogenic cycle . 100 phage is released
after lysis
6. FEATURES
At either end bears 12 nucleotides stretch with cohesive end /sticky end called Cos
sites.
It plays important role in two ways :-
Helps in recircularizations inside host cell
In lytic cycle it forms catenate i.e a linear series of replicated
DNA join together by Cos site. This site act as recognition
Genome -single linear dS DNA of about 50
kb(The genome contains 48,490 base pairs) and about
20 % is essential for exision and integration (I/E )
events so 20 Kb can be replaced by insert to be used
as vector
Has 50 gene and 5o% are needed for infection
7. λ-Bacteriophage replicates by a rolling circle mechanism
before lytic cycle and the cos sites helps in recognition by RE,
resulting in to concatameric molecules composed of several linearly
arranged recombinants. Just before packaging of the lambda DNA in
to capsid, it cleaved at cos site and a single Lambda DNA packaged
into capsid
9. Molecular aspects of Lytic cycle
• An infective lambda phage has tubular protein tail and
protein head enclosing 50 kb DNA .
• If less than 50 kb like 38 kb , the viral DNA is packed, it
becomes non infective or cannot be packed
• In contrast more than 52 Kb can not be packed.
Presence of cos allows correct packaging due to
presence of endonuclease enzymes at opening of
head.
• By assembling empty phage particle and 50 kb DNA
and tail assemblies ,infective particles can be
produced.
10. Continued…………..
• There are two Bam site that flank the I/E region
• When digested with Bam HI three sements are formed
• Left segment –contain genetic information for Head & Tail
• Right segment – for replication & Lysis
• Middle- for I/E ( Integration & Excision )
• Middle piece is replaced by DNA insert of 20 Kb
13. Process for creation of λ Vector
• Cutting of Source DNA with Bam HI
• Isolation of !5-20 Kb DNA fragments
• Ligation with the T4 Ligase of L +R and this segment
• Added Empty bacteriophage , Tail
• 50 Kb DNA is packed to form infective phage to be acted as
vector
>52 Kb <398 Kb can not be packed .
Recombinant Bacteriophage is introduced into host E, coli
14. In-vivo packaging
• In the in vivo packaging of λ DNA, first the pre-heads
are made, these preheads are the major capsid protein
encoded by gene E.
• After synthesis of preheads the single λ DNA molecules
are inserted into each pre-heads.
• These single λ DNA molecules are prepared by cutting
of concatamerized λ genomes at each cos sites.
• The maturation of preheads are done by insertion of a
minor protein named D to complete the head and the
products of other genes serve as assembly proteins,
ensuring joining of the completed tails to the
completed heads.
15. In vitro packaging
• In-vitro packaging of λ with the use of helper phage
• The in-vitro packaging of λ takes place by utilizing two E. coli strains having
λ lysogens that have several defects in the genes of pathway responsible
for packaging.
• Due to mutation in gene responsible for production of protein E, prevents
preheads being produced in strain BHB2688 (helper phage).
• In strain BHB2690 (helper phage), mutation in gene D prevent maturation
of the preheads, with packaged DNA, into complete heads.
• The functional parts of the BHB2688/BHB2690 mixed lysate having all the
components and provide all the products for correct packaging,
complementing each other’s deficiencies.
• Accordingly, recombinant λ genomes is being constructed in vitro and
enclosed into mature λ phage particles before being propagated and
replicated in host E. coli cells
17. Screening of Bacteriophage DNA
libraries
• by DNA probes
• Immunological assays
• The lytic cycle produced plaques due viral zone of lysis and
contain bacteriophage
• It is lifted on matrix and processed
• For DNA hybridization .the proteins are removed , DNA
denaured and bound to matrix . Probe of known gen is
added to identify the insert.
• Immunological assays proteins encoded by cloned gene
during lytic cycle is synthesised.These protein are
transferred along with plaque and subesquently bound to
matrix
• Positive response plaques are selected
19. Screening by Immuno assay
• The recombinant clones were filtered onto a hydrophobic
grid membrane and grown up into individual colonies, and
a replica was made onto nitrocellulose paper.
• The bacterial cells were then lysed with chloroform and the
proteins were immobilized onto the nitrocellulose paper.
• The nitrocellulose paper is then reacted with a rabbit
antibody preparation made against the particular antigenic
product to detect the recombinant clone which carries the
corresponding gene.
• The bound antibodies can be detected easily by a
colorimetric assay using goat anti-rabbit antibodies
conjugated to horseradish peroxidase.
• Positively reacting clones can be recovered from the master
hydrophobic grid membrane filter for further
characterization.