In the following slides, I have discussed the need for developing insect-resistant transgenic plants, the sources of transgenes, and methods for development
The different types of external stresses that influence the plant growth and development.
These stresses are grouped based on their characters
Biotic
Abiotic
Almost all the stresses, either directly or indirectly, lead to the production of reactive oxygen species (ROS) that create oxidative stress in plants.
This damages the cellular constituents of plants which are associated with a reduction in plant yield.
This presentation contains all the material regarding History of animal cell culture and different methods of organ and tissue culture.Hope it will be helpful..
introduction
What is virus
What is virus resistance plant
History
Gene use for develop virus resistance plant
Coat protein gene
cDNA of satellite RNA
Defective viral genome
Antisense RNA approach and
Ribozyme – mediated protection
conclusion
References
Presented by- MD JAKIR HOSSAIN
Doctoral Research Scholar
Department of Agricultural Genetic Engineering ,
Faculty of Agricultural Sciences and Technologies,
Nigde Omer Halisdemir University, Turkey
E. Mail- mjakirbotru@gmail.com
The different types of external stresses that influence the plant growth and development.
These stresses are grouped based on their characters
Biotic
Abiotic
Almost all the stresses, either directly or indirectly, lead to the production of reactive oxygen species (ROS) that create oxidative stress in plants.
This damages the cellular constituents of plants which are associated with a reduction in plant yield.
This presentation contains all the material regarding History of animal cell culture and different methods of organ and tissue culture.Hope it will be helpful..
introduction
What is virus
What is virus resistance plant
History
Gene use for develop virus resistance plant
Coat protein gene
cDNA of satellite RNA
Defective viral genome
Antisense RNA approach and
Ribozyme – mediated protection
conclusion
References
Presented by- MD JAKIR HOSSAIN
Doctoral Research Scholar
Department of Agricultural Genetic Engineering ,
Faculty of Agricultural Sciences and Technologies,
Nigde Omer Halisdemir University, Turkey
E. Mail- mjakirbotru@gmail.com
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.
Introduction
Definition of an Insect Resistant Plant
What is the Bt gene?
History
The crystal ( cry)Proteins
Definition of cry protein
How does Bt work?
Mechanism of Bt toxicity
Mode of Action of Insecticidal Crystal Protein
Bt Technology
The Insect Resistance Problem
Advantages
Limitations
Conclusion
References
Scale up means increasing the quantity or volume of cell culture. For animal cells, the scale up strategies are dependent upon cell types or i.e. whether the cells requires matrix for attachment and growth ( adherent cell culture) or grows freely in suspended form in aqueous media. The scaling up principle for adherent cells are just to increase surface area for attachment while for suspension culture is to increase culture volume. This presentation enlightens the reader about different methods of scaling up of cells culture. Readers are also provided with sample questions for better understanding
Genetic manipulation of plant and animal cells have to be confirmed for further application. One such confirmatory method is the use of stains/dyes which produces fluorescence when the recombination is successful.
In nuclear biology and molecular biology, a marker gene is a gene used to determine if a nucleic acid sequence has been successfully inserted into an organism's DNA.
The direct microinjection of DNA into the cytoplasm or nuclei of cultured cells is sometimes used as a transfection method. It is highly efficient at the level of individual cells. The most significant use of this technique is introduction of DNA into the oocytes, eggs and embryos of animals, either for transient expression analysis (e.g. in fish or Xenopus) or to generate transgenic animals (e.g. mice, Drosophilathis). The procedure is time consuming and only a small number of cells can be treated. Originally, this technique was used for the transformation of cells that were resistant to any other method of transfection. Stable transfection efficiencies are extremely high, in the order of 20%, and very small quantities of DNA are sufficient.
This technique provides direct nuclear delivery of DNA avoiding the endogenous pathway and also ensures that the DNA is delivered intact. Microinjection is suitable for the introduction of large vectors such as YACs into the pronuclei of fertilized mouse eggs. DNA delivered in this manner must be very pure so it needs a lot of preparation as it is necessary to avoid fragmentation. Shearing can also occur in the delivery needle, and large DNA fragments are often protected by suspension in a high salt buffer and/or mixing with polyamines and other protective agents. Now transfection of cultured cells is automated with computer-controlled micromanipulation and microinjection processes as well as the automated production of injection capillaries and the standardization of cell preparation procedure.
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.
Introduction
Definition of an Insect Resistant Plant
What is the Bt gene?
History
The crystal ( cry)Proteins
Definition of cry protein
How does Bt work?
Mechanism of Bt toxicity
Mode of Action of Insecticidal Crystal Protein
Bt Technology
The Insect Resistance Problem
Advantages
Limitations
Conclusion
References
Scale up means increasing the quantity or volume of cell culture. For animal cells, the scale up strategies are dependent upon cell types or i.e. whether the cells requires matrix for attachment and growth ( adherent cell culture) or grows freely in suspended form in aqueous media. The scaling up principle for adherent cells are just to increase surface area for attachment while for suspension culture is to increase culture volume. This presentation enlightens the reader about different methods of scaling up of cells culture. Readers are also provided with sample questions for better understanding
Genetic manipulation of plant and animal cells have to be confirmed for further application. One such confirmatory method is the use of stains/dyes which produces fluorescence when the recombination is successful.
In nuclear biology and molecular biology, a marker gene is a gene used to determine if a nucleic acid sequence has been successfully inserted into an organism's DNA.
The direct microinjection of DNA into the cytoplasm or nuclei of cultured cells is sometimes used as a transfection method. It is highly efficient at the level of individual cells. The most significant use of this technique is introduction of DNA into the oocytes, eggs and embryos of animals, either for transient expression analysis (e.g. in fish or Xenopus) or to generate transgenic animals (e.g. mice, Drosophilathis). The procedure is time consuming and only a small number of cells can be treated. Originally, this technique was used for the transformation of cells that were resistant to any other method of transfection. Stable transfection efficiencies are extremely high, in the order of 20%, and very small quantities of DNA are sufficient.
This technique provides direct nuclear delivery of DNA avoiding the endogenous pathway and also ensures that the DNA is delivered intact. Microinjection is suitable for the introduction of large vectors such as YACs into the pronuclei of fertilized mouse eggs. DNA delivered in this manner must be very pure so it needs a lot of preparation as it is necessary to avoid fragmentation. Shearing can also occur in the delivery needle, and large DNA fragments are often protected by suspension in a high salt buffer and/or mixing with polyamines and other protective agents. Now transfection of cultured cells is automated with computer-controlled micromanipulation and microinjection processes as well as the automated production of injection capillaries and the standardization of cell preparation procedure.
Mechanism of insect resistance in plants (non preference, antibiosis, tolerance and avoidance) – nature of insect resistance – genetics of insect resistance – horizontal and vertical – genetics of resistance – sources of insect resistance – breeding methods for insect resistance – problems in breeding for insect resistance – achievements.
WHAT IS VACCINE
PROPERTIES OF IDEAL VACCINE
TYPES OF VACCINEs
TRADIONTIONAL VS EDIBLE VACCINES
EDIBLE VACCINES :- INTRO AND DEFINITION
STANDARDS FOR EDIBLE VACCINE
HISTORY OF EDIBLE VACCINE
WHY TO CHOOSE EDIBLE VACCINE?
CRITERIA FOR HOST PLANT
DEVELOPING AN EDIBLE VACCINE
METHOD OF VACCINE PRODUCTION
HOW TO MAKE EDIBLE VACCINE
HOW EDIBLE VACCINE WORK (MECHANISM)
FACTOR AFFECTING EDIBLE VACCINE
PROS OF EDIBLE VACCINE
CONS OF EDIBLE VACCINE
PLANTS USED FOR EDIBLE VACCINE PRODUCTION
PROS AND CONS OF SELECTED HOST PLANT
APPLICATION
FUTURE PROSPECTS
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
2. z
INTRODUCTION
Insects are the most diverse species of animals that compete for food, fibre and natural
resources.
One-fifth of the world's total crop production is destroyed by them.
Indiscriminate use of chemical pesticides results in
Environmental pollution
Development of resistance
Killing of useful insects
Adverse effect in human health
Resistant crop varieties are developed using conventional breeding methods, but this
approach is considered inefficient because the methods depend on germplasm and are
time and resource-consuming.
3. z
TO OVERCOME THESE PROBLEMS
Transgenic crops were generated by introducing the specific gene
of interest into the host with the help of genetic engineering
technology. Expression of these genes coding for insecticidal
activity enables the development of "INSECT RESISTANT
TRANSGENIC PLANTS."
The first transgenic insect-resistant plant was produces by
introducing cry gene from Bacillus thuringiensis bacterium. Later
the cry gene containing transgenic maize and cotton was
produced commercially.
7. z
Mechanism Of
Action
cry gene- These toxins will bind to the specific
receptors in the gut of an insect and are solubilized
and activated by specific proteinases, induce the
formation of pores in the insect’s midgut epithelial
membrane, resulting in cell lysis and death.
vip gene- It is responsible for disruption and
swelling of midgut epithelial cells of insects
by osmotic lysis.
choM- Cholesterol oxidase kills the larvae of
boll weevil by disrupting the midgut epithelial
cells at lower doses, and at higher doses cells
are lysed.
8. z
Chitinase- Makes insect pests more
vulnerable to attack by different
pathogens.
Avidin- binds with high affinity to a
ligand biotin that is required by all life
forms as a coenzyme.
Mechanism Of
Action
9. z Lectins- Carbohydrate binding proteins which
disrupt the function by binding to the
midgut epithelial cells of an insect. Thus
inhibitting absorption of nutrients.
Proteinase inhibitors- proteinase inhibitors
which affects the digestion of protein in the gut of
an insect and leads to the deficiency of essential
amino acids.
α-Amylase inhibitors- have a role in the
defense mechanism of plants against insects
by forming a complex with amylases of certain
insect species
Mechanism Of
Action
11. z
Common Methods For
Gene Delivery
Electroporation
Microinjection
Polyethylene Glycol
Biolistics
Agrobacterium
12. z
Tactics For
Deployment
Transgenic plants need to
be integrated with pest
management strategies
as insects have a capacity
to develop resistance to a
wide array of insecticides
• Single Gene
• Multiple gene(Gene Pyramiding)
• Chimeric gene
Gene Strategies
• Constitutive
• Tissue specific
• Inducible(ex. Wounding)
Gene Promoter
• High Dose
• Low dose
• Mixture
Gene Expression
• Uniform single gene
• Mixture of genes
• Gene rotation
• Mosaic Planting
• Refuges(spatial/temporal)
Field Tactics
13. z
Conclusion And Perspective
The use of transgenic plants provides a way to increase crop production,
benefiting farmers in developed and developing countries by using the resistant
genes either singly or in combination.
Continuing research on new sources of resistance is essential for the long term
control of insect pests.
Studies on the expression and potential of the new insecticidal genes can be
performed in model plants such as tobacco and Arabidopsis thaliana. Then, the
selected gene(s) must be introduced into the target crop.
The introduction of insect-resistant crops must be accompanied by a resistance
management strategy.
14. z
References
1. Insect-Resistant Plants
D. Easwar Rao, K. Divya, I.V.S.N. Prathyusha, Ch. Rama Krishna, K.V. Chaitanya*
GITAM UNIVERSITY, VISAKHAPATNAM, INDIA
2. Biotechnological Prospects for Engineering
Insect-Resistant Plants1[C]
John A. Gatehouse*
School of Biological and Biomedical Sciences, Durham University, Durham DH1 3LE,
United Kingdom
3. Insect-Resistant Plants
D. Easwar Rao, K. Divya, I.V.S.N. Prathyusha, Ch. Rama Krishna,
K.V. Chaitanya*
GITAM UNIVERSITY, VISAKHAPATNAM, INDIA