The document describes the phage display technique, which involves genetically fusing a protein to the coat protein of a bacteriophage virus. This allows for the display of large protein and peptide libraries on the surface of the phage. The technique has various applications, including epitope determination, enzyme engineering, organ targeting, and isolating allergens. It involves creating a vector fusion, binding and selecting phages to a target, washing, eluting bound phages, and amplifying the process through bacterial infection.
The three hybrid system of yeast has been described in this ppt. Yeast one Hybrid system, yeast two hybrid system and yeast 3 hybrid system is explained. This explain about the DNA-protein interaction and Protein-DNA-Protein interaction.
The three hybrid system of yeast has been described in this ppt. Yeast one Hybrid system, yeast two hybrid system and yeast 3 hybrid system is explained. This explain about the DNA-protein interaction and Protein-DNA-Protein interaction.
This is technique used widely for protein separation from a mixture and is very easy and less costly method. Slides cover all essential points about EMSA and it is quite interesting to know that how it detect and separate different proteins and their mobility shift assay.
Genome editing is one of the most important tools which supports genetic engineering. It is based on the naturally occurring mechanism of DNA recombination which involves the initiation of breaks with the double stranded DNA followed by repair by the endogenous DNA polymerases.
Conventional techniques such as gene knockouts using P-elements and transposable genetic elements have been superseded by more accurate genome editing methods such as TALENs and CRISPR/Cas.
Expression and purification of recombinant proteins in Bacterial and yeast sy...Shreya Feliz
This presentation gives the information about bacterial and yeast system as host for expressing recombinant proteins, suitable vectors, strains of host, Pros and cons of this system, different purification techniques and commercially available proteins produced so far by this system.
Cell culture based vaccine??
Cell cultures involve growing cells in a culture dish, often with a supportive growth medium. A primary cell culture consists of cells taken directly from living tissue, and may contain multiple types of cells such as fibroblasts, epithelial, and endothelial cells.
In the United States, 10 different vaccines for chicken pox, hepatitis A, polio, rabies, and rubella are cultured on aborted tissue from two fetal cell lines known as WI-38 and MRC-5. These vaccines are chicken pox, hep-A, hep-A, hep-A/hep-B, polio, rabies, rubella, measles/rubella, mumps/rubella, and MMR II (measles/mumps/rubella).
Transfection methods (DNA to host cell) Erin Davis
Transfection of DNA to host cell can be done by various methods in lab scale.Gene gun,electroporation,lipofection .These methods are used to transfer DNA to the host cell.
This is technique used widely for protein separation from a mixture and is very easy and less costly method. Slides cover all essential points about EMSA and it is quite interesting to know that how it detect and separate different proteins and their mobility shift assay.
Genome editing is one of the most important tools which supports genetic engineering. It is based on the naturally occurring mechanism of DNA recombination which involves the initiation of breaks with the double stranded DNA followed by repair by the endogenous DNA polymerases.
Conventional techniques such as gene knockouts using P-elements and transposable genetic elements have been superseded by more accurate genome editing methods such as TALENs and CRISPR/Cas.
Expression and purification of recombinant proteins in Bacterial and yeast sy...Shreya Feliz
This presentation gives the information about bacterial and yeast system as host for expressing recombinant proteins, suitable vectors, strains of host, Pros and cons of this system, different purification techniques and commercially available proteins produced so far by this system.
Cell culture based vaccine??
Cell cultures involve growing cells in a culture dish, often with a supportive growth medium. A primary cell culture consists of cells taken directly from living tissue, and may contain multiple types of cells such as fibroblasts, epithelial, and endothelial cells.
In the United States, 10 different vaccines for chicken pox, hepatitis A, polio, rabies, and rubella are cultured on aborted tissue from two fetal cell lines known as WI-38 and MRC-5. These vaccines are chicken pox, hep-A, hep-A, hep-A/hep-B, polio, rabies, rubella, measles/rubella, mumps/rubella, and MMR II (measles/mumps/rubella).
Transfection methods (DNA to host cell) Erin Davis
Transfection of DNA to host cell can be done by various methods in lab scale.Gene gun,electroporation,lipofection .These methods are used to transfer DNA to the host cell.
New Progress in Pyrosequencing for Automated Quantitative Analysis of Bi- or ...QIAGEN
Pyrosequencing is a highly flexible technology based on sequencing-by-synthesis for the rapid and quantitative analysis of any type of sequence variation. The real-time output delivers high-resolution sequence information, making pyrosequencing highly suitable for applications ranging from biallelic or multiallelic SNP analysis, DNA methylation quantification to complex mutation analysis of multiple sequence variations in a single run.
In this slideeck, we introduce the new PyroMark Q48 Autoprep system which enables fully automated template preparation integrated in the pyrosequencing workflow. In addition, a new Multiple Primer Dispensation (MPD) strategy is presented which allows fully automated dispensation of sequencing primer, offering a seamless workflow from samples to quantitative genotyping results.
This slidedeck focuses on the following topics
• Pyrosequencing technology and workflow in genotyping analysis
• Introduction into the new PyroMark Q48 Autoprep
• MPD strategy for a seamless automated pyrosequencing workflow
Join us and learn how you can apply the new pyrosequencing system and protocol to your variant analysis or genotyping research
Analysis of Peroxisomal Lipid Metabolism in the Oleaginous Microalga Nannochloropsis and Development of Synthetic Biology Tools for Genetic Engineering
it will help you to understand how the protein microarrays are made, what are the different types and what all purposes they are used for. its very useful ppt
New Progress in Pyrosequencing for DNA MethylationQIAGEN
Pyrosequencing is a highly flexible technology that lets you rapidly analyze short- to medium-length sequences fast and quantitatively with high accuracy. The real-time, high-resolution sequence output makes the technology highly suitable for applications including complex mutation analysis, microbial identification and DNA methylation quantification.
The main bottleneck in Pyrosequencing has been limited sequence length, which is critical for some applications. Our new technology, software, and chemistry overcome this bottleneck and give sequence reads that are typically twice as long as those from previous PyroMark systems. The new PyroMark Q24 Advanced system also reduces background noise, improving quantification even at sites distant from the sequencing start. The new system is ideal for applications requiring analysis of longer sequences, such as DNA methylation analysis in epigenetic research, frequency determination in mutation analysis, and various de novo sequencing applications.
In this presentation, we will discuss the following applications and technology improvements:
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Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
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.
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.
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
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. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
INTRODUCTION
HISTORY
TECHNIQUE
•CREATION OF VECTOR AND FUSION
•BINDING AND SELECTION
•WASHING
•ELUTION
•AMPLIFICATION
APPLICATION
•EPITOPE DETERMINATION
•ENZYMATIC APPLICATION
•ORGAN TARGETING
•GENE DELIVERY
•ISOLATION OF ALLERGENS
ADVANTAGES
DISADVANTAGES
CONCLUSION
SUMMARY
REFERENCES
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3. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
Bacteriophages are viruses that infect bacterial cells.
Infected cells are used as a host to replicate viruses.
It is an in vitro selection technique using a protein
genetically fused to the coat protein of a bacteriophage.
E. coli phages used due to ease of culture and quick
regeneration.
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4. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
George P. Smith 1985 first introduced Phage
display technology
Bass et al 1990 A large number of
phage displayed
peptide and protein
libraries have been
constructed.
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5. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
This technique is called phage display because it
involves the display of protein on the surface of
Bacteriophage.
In this , cloning vector used for phage display.
Following steps involved in the phage display-
•Creation of vector and fusion
•Binding and selection
•Washing
•Elution
•Amplification
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6. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
Creation of vector and fusion
• Recombinant DNA technology to incorporate
foreign cDNA of interest into viral DNA.
• The cloned gene becoming fused with a gene for
the phage coat protein.
• So the protein will be displayed on outside of
phage particles.
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7. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
Binding and selection
• Can apply standard affinity techniques to
capture phage by taking advantage of
displayed proteins.
• Pass solutions of amplified phages over solid
support with antigens or receptors bound to it.
• Phages with affinity to support bind.
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8. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
Washing
• Unbound phages are washed away leaving only
those showing affinity for the receptors.
Elution
• Bound phages can be eluted by disrupting the
protein bonding interactions.
• Acidic buffers, Alkaline buffers, Urea, addition of
soluble ligend for receptor.
• Can also add host cells to infect.
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9. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
Amplification
• Eluted phages showing specificity are used
to infect new host cells for amplification.
• Cycle repeated 2-3 times for stepwise selection of
best binding sequence.
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10. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
Fig. No.- 1, Steps involved in phage display technique.
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12. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
Phage display technique have following
application-
•EPITOPE DETERMINATION
•ENZYMATIC APPLICATION
•ORGAN TARGETING
•GENE DELIVERY
•ISOLATION OF ALLERGENS
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13. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
Epitope determination
•Peptide libraries have been used to determine the
epitope to which an antibody binds.
•Therefore, it is possible to define the region of a
protein recognized by an antibody.
• Phage libraries were used to define peptide
structures recognized by major histocompatibility
(MHC) molecules.
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14. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
ENZYMATIC APPLICATION
•In enzymology, it has been applied for mechanistic-
based studies and to generate enzyme variants with
new or improved properties.
• Enzymes with a broader range of substrates, for
instance, would have improved function and allow
certain advantages to its host.
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15. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
ORGAN TARGETING
•In this technique phage libraries are injected
intravenously into animals and then organs or
tissues are collected and examined for phage bound
to tissue-specific endothelial cell markers.
GENE DELIVERY
•Gene delivery to mammalian cells has also been
accomplished by the use of single and double-
stranded phage.
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16. ISOLATION OF ALLERGENS
•Fusion proteins created by the principle of linking
the phage phenotype to its genetic information
are covalently associated with the phage particle.
•Therefore, cDNA libraries displayed on phage
surface can be screened for the presence of
specific clones by affinity purification.
PHAGE DISPLAY-TECHNIQUE AND APPLICATION
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17. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
•Phage clones that bind to IgE may be selected by
screening and enrichment of phage libraries
against serum IgE immobilized in a solid phase.
•The amino acid sequence of surface-expressed
allergens can be clarified by sequencing the DNA
of the integrated section of the phage, as there is
a physical linkage between its genotype and
phenotype.
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•Easy to screen large of clones >109.
• Easy to amplify selected phages in E. coli.
•Selection process easy and already in use in various
forms.
•Can create Phage library variation by inducing
mutations, using error prone PCR, etc.
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•Might not have long enough peptide insert so
critical folding can be disrupted.
•Could lose phage variations if first bind/wash
step too stringent
•Affinities or binding that results during
selection might not work in vivo.
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20. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
Phage display allows the presentation of large peptide
and protein libraries on the surface of filamentous
phage, which leads to the selection of peptides and
proteins, including antibodies, with high affinity and
specificity to almost any target.
The technology involves the introduction of exogenous
peptide sequences into a location in the genome of the
phage capsid proteins.
The encoded peptides are expressed or "displayed" on
the phage surface as a fusion product with one of the
phage coat proteins.
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21. PHAGE DISPLAY-TECHNIQUE AND APPLICATION
Bacteriophages are viruses that infect bacterial cells.
Infected cells are used as a host to replicate viruses.
Phage display technique involves the following steps-
•Creation of vector and fusion
•Binding and selection
•Washing
•Elution
•Amplification
Phage display technique have following
application-
•Epitope determination
•Enzymatic application
•Organ targeting
•Gene delivery
•Isolation of allergens
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T. A. Brown 2008 Gene cloning and DNA
analysis
Some contents from net-
www.mupnet.com/jocm.htm
www.bio.anl.gov/antibody.html
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