CaMV Genome organization & their replication, Cauliflower Mosaic Virus belong to Group VII (ds-DNA-RT), Open circular double stranded DNA of 80kb and CaMV replicates by reverse transcription
inroduction:Plant viruses are viruses that affect plants.
Pathogenic to higher plants.
. Harmless to human and other animals.
Reduce plant crop yield and quality of crops.
Some may be able to multiply within the bodies
Of aphids and nematodes.
History:Beijernick ( 1897) coined the latin name “VIRUS” meaning Poison. He studied plant juices and found they caused healthy plants to become sick.
Wendell Stanley (1935) crystallized sap from sick Tobacco plants. He discovered viruses were made of nucleic acids and proteins.
Geminivirus:one of the family of plant virus.
Currently over 360 species in this family, divided among 9 genera.
Diseases associated with this family include bright yellow mosaic , yellow mosaic, yellow mottle, leaf curling, stunting, streaks, reduced yields.
Ss circular dna diverge in both directions from a virion strand origin of replication (AMBISENSE).
Virus Classification:Group – Group II (ssDNA)
Order - Unassigned
Family - Geminiviridae
Genera – Becurtovirus Grablovirus
Begomovirus Mastrevirus
Capulavirus Topocuvirus
curtovirus Turncurtovirus
Eragrovirus
Structure: have Circular single-stranded DNA.
Genome is either in two segments.
The non-segmented genome is 2500-3000 nucleotides long, and the segmented genome is 4800-5600 nucleotides long.
The genome encodes for both structural and non-structural proteins.
In geminivirus, both segments must be transmitted to the host for a full systemic infection to occur.
Virion Sturcture:Geminivirus are non-enveloped, icosahedral virions that consists of a capsid.
The capsid is germinate, or twinned, and consists of 22 Capsomers.
The capsid is 30nm long and has a diameter of 18-20nm.
Symptoms:the time of infection, the virus strains and the presence of mixed infections.
Common symptoms are stunting, curling, and twisting of leaves.
Short internodes and stunted appearance , no apical growth caused by early infection.
Replication:Geminivirus encodes only a few proteins, thus they need to dependent host cell factors for replication.
These factors are DNA polymerase and repair polymerase to amplify their genome.
Replicate by a rolling circle mechanism like bacteriophages such as M13, and many plasmids.
CaMV Genome organization & their replication, Cauliflower Mosaic Virus belong to Group VII (ds-DNA-RT), Open circular double stranded DNA of 80kb and CaMV replicates by reverse transcription
inroduction:Plant viruses are viruses that affect plants.
Pathogenic to higher plants.
. Harmless to human and other animals.
Reduce plant crop yield and quality of crops.
Some may be able to multiply within the bodies
Of aphids and nematodes.
History:Beijernick ( 1897) coined the latin name “VIRUS” meaning Poison. He studied plant juices and found they caused healthy plants to become sick.
Wendell Stanley (1935) crystallized sap from sick Tobacco plants. He discovered viruses were made of nucleic acids and proteins.
Geminivirus:one of the family of plant virus.
Currently over 360 species in this family, divided among 9 genera.
Diseases associated with this family include bright yellow mosaic , yellow mosaic, yellow mottle, leaf curling, stunting, streaks, reduced yields.
Ss circular dna diverge in both directions from a virion strand origin of replication (AMBISENSE).
Virus Classification:Group – Group II (ssDNA)
Order - Unassigned
Family - Geminiviridae
Genera – Becurtovirus Grablovirus
Begomovirus Mastrevirus
Capulavirus Topocuvirus
curtovirus Turncurtovirus
Eragrovirus
Structure: have Circular single-stranded DNA.
Genome is either in two segments.
The non-segmented genome is 2500-3000 nucleotides long, and the segmented genome is 4800-5600 nucleotides long.
The genome encodes for both structural and non-structural proteins.
In geminivirus, both segments must be transmitted to the host for a full systemic infection to occur.
Virion Sturcture:Geminivirus are non-enveloped, icosahedral virions that consists of a capsid.
The capsid is germinate, or twinned, and consists of 22 Capsomers.
The capsid is 30nm long and has a diameter of 18-20nm.
Symptoms:the time of infection, the virus strains and the presence of mixed infections.
Common symptoms are stunting, curling, and twisting of leaves.
Short internodes and stunted appearance , no apical growth caused by early infection.
Replication:Geminivirus encodes only a few proteins, thus they need to dependent host cell factors for replication.
These factors are DNA polymerase and repair polymerase to amplify their genome.
Replicate by a rolling circle mechanism like bacteriophages such as M13, and many plasmids.
vaccine is a biological preparation that provides active acquired immunity to a particular disease. A vaccine typically contains an agent that resembles a disease-causing microorganism and is often made from weakened or killed forms of the microbe, its toxins, or one of its surface proteins. The agent stimulates the body's immune system to recognize the agent as a threat, destroy it, and to further recognize and destroy any of the microorganisms associated with that agent that it may encounter in the future.
HISTORY OF VACCINES-
EDWARD JENNER conduct experiments in 1796 that lead to the creation of the first smallpox vaccine for prevention of smallpox.
A vaccine for RABIES is developed by LOUIS PASTEUR .
Vaccine for COLERA and TYPHOID were developed in 1896 and PLAGE vaccine in 1887.
The first DIPHTHERIA vaccine is developed in about 1913 by EMIL ADOLPH BEHRING,WILLIAM HALLOCK PARK.
The whole cell PERTUSIS vaccines are developed in 1914.
A TETANUS vaccine is developed in 1927.
in this chapter covers the symptoms modulation and diseases severity increases and decreases. and role of SiRNA in diseases severity reduction. and also covers the types of SRNA..
Topics included - Introduction; explanation; examples like blue white screening method, antibiotic resistance; Extra information regarding - detection of oncogene in vertebrates and role of sleeping beauty; Merits and demerits of insertional inactivation.
Bacteriophage- types, structure and morphology of t4 phage, morphogenesisDr. Dinesh C. Sharma
Escherichia virus T4 is a species of bacteriophages that infect Escherichia coli bacteria. It is a member of virus subfamily Tevenvirinae (not to be confused with T-even bacteriophages, which is an alternate name of the species). T4 is capable of undergoing only a lytic lifecycle and not the lysogenic lifecycle.
Synopsis
Introduction
History
Definition
Need for edible vaccine
Plants normally used for production of
edible vaccine
Production
Mode of application
Advantages
Disadvantages
Application
Conclusion
References
To decrease our world hunger and to make the plant more nutritious the transgenic technique was developed. This the basis of the transgenic plant and its technique
vaccine is a biological preparation that provides active acquired immunity to a particular disease. A vaccine typically contains an agent that resembles a disease-causing microorganism and is often made from weakened or killed forms of the microbe, its toxins, or one of its surface proteins. The agent stimulates the body's immune system to recognize the agent as a threat, destroy it, and to further recognize and destroy any of the microorganisms associated with that agent that it may encounter in the future.
HISTORY OF VACCINES-
EDWARD JENNER conduct experiments in 1796 that lead to the creation of the first smallpox vaccine for prevention of smallpox.
A vaccine for RABIES is developed by LOUIS PASTEUR .
Vaccine for COLERA and TYPHOID were developed in 1896 and PLAGE vaccine in 1887.
The first DIPHTHERIA vaccine is developed in about 1913 by EMIL ADOLPH BEHRING,WILLIAM HALLOCK PARK.
The whole cell PERTUSIS vaccines are developed in 1914.
A TETANUS vaccine is developed in 1927.
in this chapter covers the symptoms modulation and diseases severity increases and decreases. and role of SiRNA in diseases severity reduction. and also covers the types of SRNA..
Topics included - Introduction; explanation; examples like blue white screening method, antibiotic resistance; Extra information regarding - detection of oncogene in vertebrates and role of sleeping beauty; Merits and demerits of insertional inactivation.
Bacteriophage- types, structure and morphology of t4 phage, morphogenesisDr. Dinesh C. Sharma
Escherichia virus T4 is a species of bacteriophages that infect Escherichia coli bacteria. It is a member of virus subfamily Tevenvirinae (not to be confused with T-even bacteriophages, which is an alternate name of the species). T4 is capable of undergoing only a lytic lifecycle and not the lysogenic lifecycle.
Synopsis
Introduction
History
Definition
Need for edible vaccine
Plants normally used for production of
edible vaccine
Production
Mode of application
Advantages
Disadvantages
Application
Conclusion
References
To decrease our world hunger and to make the plant more nutritious the transgenic technique was developed. This the basis of the transgenic plant and its technique
Introduction: Biotechnology is an emerging field of research as it has the potential to solve many biological problems which could not be solved till now with conventional techniques.
The use of biology to develop technologies and products for the welfare of human beings is known as Biotechnology. It has various applications in different fields such as Therapeutics, Diagnostics, Processed Food, Waste Management, Energy Production, Genetically Modified Crops etc.
Biotechnology means 'applications of scientific and engineering principles to biological processes to provide goods and services'. Full understanding of biological processes is possible with detailed analysis of gene structure and function i.e. the Genetic Engineering means the introduction of manipulated genetic material (DNA) into a cell in such a way as to replicate and be passed on to progeny cells'. The outcome is attractive and promising.
Genetic modification of plants involves adding a specific stretch of DNA into the plant's genome, giving it new or different characteristics.
INTRODUCTION
EXAMPLES
CONCLUSION
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.
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.
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 .
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.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
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.
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.
4. TRANSGENIC PLANTS
DNA is modified using Genetic Engineering
techniques.
Transgenic Plant contains Genes inserted Artificially
called Transgenes.
Provides advantages in producing plants wich are
Drought Resistant,cold tolerance, pest resistant, etc…
5. Methods Employed for
Transformation
Direct gene Transfer-Electroporation,Particle
Bombardment,Micro injection,PEG mediated.
In-direct gene Transfer-Virus mediated,Agrobacterium
mediated
Agrobacterium mediated transformation is the only
high Efficient Transformation Till date
than,Remaining Transformation Techniques.
8. RICE
Rice,wheat,maize are worlds 3 leading food crops
Rice is staple food of more than half of the worlds
population
More than 3.5 billion people depend on rice
9. Challenges in Rice
Disease,many disease such as bacterial,fungal,viral
diseases,pests
Create huge loss to farmers
Some fungal disease like Blast created Famines.like the
great bengal famine
Rice requires huge water for cultivation
Rice is not a salt tolerant plant.
Rice is not a Drought Resistant crop.
11. TRANSGENIC RICE
Pid2-Rice plants
resistant to Rice blast
Cry1la5-rice resistant to
stemborrer
Am sod-rice plant
showing high level salt
tolerance
AtDREBIA-plants
showing improved
drought tolerance
12. Agb,mediatedTransformation in
Rice
First step is to Transform
the Agrobacterium by
using molecular biology
tools
Inserting our GOI with
Marker,into the bacteria.
Once Agrobacterium is
engineered it can be
multiplied easily.
Agrobacterium with
GOI,is streaked on LB
plates.
Agb,is selected with
R,K,T,S
LB plates are stored at 4
0 c .
13. EXPLANT
Rice is a monocot plant.
Monocot plants are
complicated compared
to dicots
Unlike other
Dicots,Explants used in
monocots are Immature
Embryos or Calli.
14. Immature rice for transformation
Immature embryos are
used as Explants in rice
transformation.
Rice is grown in a green
house for obtaining good
immature embryos at the
right stage.
15. Sterilization and isolation of
embryos
Immature rice is de-
husked,surface sterilized
with 2% Naocl,for 20
mins.
Using a
microscope,immature
embryos are dissected.
Embryos are collected in
liquid media (infection
media)in ependorf tubes
16. Transformation
All the Embryos are
collected in liquid
media.
Collected embryos are
pre treated,that means
they are kept in water
bath at 430Cfor 30
mins,and ice shock for 1
min.
Agrobacterium culture is
freashly streaked on LB
plates,one day before
Transformation.
O.D for infection should
be set as desired.
Desired OD is adjusted
and culture is
resuspended in liquid
infection media.
17. Infection and co-cultivation
Embryos after pre
treatment are infected with
agrobacterium suspended
media for 15 mins
Agrobacterium suspension
is removed,and embryos
are kept in co-cultivation
media,in dark for 7 days.
This step is called co-
cultivation.
18. RESTING AND SELECTION
The embryos are kept in resting
media,ie..media can kill
agrobacterium but withou any
selection pressure.so embryos
can grow(both transformed and
non-transformed).
After Resting,embryos are
moved to Selection
Media.ie..media with
hygromycin or kanamycin…
Selection media is that in wich
only Transformed cells survive
and remaining cells die,so
Transformed embryo forming
calli survive and Remaining Die.
19. REGENERATION
After 3 rounds of
selection,calli formed is
kept for Regeneration of
transformed plant.
After 3 regeneration
subcultures on suitable
media,Plants are
Regenerated on media.
Regenerated plants are
Transferred to Rooting
media.
After rooting plants are
transffered to pots for
whole plant Regeneration.
20. GUS ASSAY
GUS gene is used as Reporter
gene in Transformation
studies
The enzyme β-glucuronidase
cleaves the substrate X-gluc
into glucuronic acid and
indoxyl derivative. The
indoxyl derivative dimerizes
and is then oxidized to form
insoluble, dark indigo dye the
presence of which can
indicate the site of expression
of the gene
21. Geno typing of Transformed Plants
Complete regenerated
plants in a green house
can be analyzed for
GOI(gene of interest),via
pcr, southern blot or
other molecular
techniques.
PCR,GUS(if used in gene
construct) can
demonstrate if Plant is
Transformed
