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.
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.
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.
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.
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.
Introduction
Components of binary vector
Development of binary vector system
Properties of binary vector
Types of binary vector
Plant transformation using binary vector
Advantage of using binary vector
Conclusion
References
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
MBB 501 PLANT BIOTECHNOLOGY
INFORMATION ABOUT DIFFERENT DNA MODIFYING ENZYMES
WHAT IS AN ENZYME?
Alkaline Phosphatase
Polynucleotide kinase
Terminal deoxyneucleotidyl transferase
Nucleases
Exonuclease
Bal31 Exonuclease III
Endonuclease
S1 endonulease
Deoxyribonuclease 1 (Dnase 1)
RNase A
RNase H
Restriction Endonuclease
PvuI
PvuII
Different types of endonuclease enzymes
The recognition sequences for some of the most frequently used restriction endonucleases.
Categorization of enzymes
Isoschizomers
Neoschizomers
Isocaudomers
Introduction
Components of binary vector
Development of binary vector system
Properties of binary vector
Types of binary vector
Plant transformation using binary vector
Advantage of using binary vector
Conclusion
References
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
MBB 501 PLANT BIOTECHNOLOGY
INFORMATION ABOUT DIFFERENT DNA MODIFYING ENZYMES
WHAT IS AN ENZYME?
Alkaline Phosphatase
Polynucleotide kinase
Terminal deoxyneucleotidyl transferase
Nucleases
Exonuclease
Bal31 Exonuclease III
Endonuclease
S1 endonulease
Deoxyribonuclease 1 (Dnase 1)
RNase A
RNase H
Restriction Endonuclease
PvuI
PvuII
Different types of endonuclease enzymes
The recognition sequences for some of the most frequently used restriction endonucleases.
Categorization of enzymes
Isoschizomers
Neoschizomers
Isocaudomers
this presentation is about reporter gene essay, its types, blue white screening and its application, Antibiotic resistance gene and Herbicide resistance markers
Use of reporter genes in the process of selection of the transformants from the non transformants, and the current use of these reporter genes as the Desired genes.
A transplastomic plant is a genetically modified plant in which the new genes have not been inserted in the nuclear DNA but in the DNA of the chloroplasts.
The presentation describes the advantages of plastid transformation over 'conventional' nuclear transformation, hurdles to plastid transformation, its advantages. The presentation also covers some successful plastid engineering and its potential.
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.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
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.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
(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.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
2. What is a marker?
&
What is the purpose of the
marker?
Answer: it is a molecule, which is used to identify or
tag a cell/group of cells.
3. Introduction
Marker genes are used as tools in
o molecular biology (transformation studies)
o metagenomics (study direct environmental sample)
o phylogenetic analysis (genetic lineage)
They are used to determine whether a particular piece of DNA
or gene of interest is successfully inserted into the target DNA.
It is also used to determine orthologous gene groups.
It is present next to the transgene.
It is of two types: screenable markers and selectable markers.
4. Screenable Markers
Screenable markers are also referred to as reporter genes or
scorable markers.
Often accompanied with colour change or visible phenotypic
changes in the tissues of transformed organisms.
They do not affect the growth of the cells.
Preferred when a large group of cells have to be screened.
They are used for quantitative assays.
5. Types of Screenable Markers
Green Fluorescent Protein (GFP)
o The gene is derived from jelly fish (Aequorea victoria).
o This protein has 238 amino acids of which amino acids serine, tyrosine and glycine
imparts the chromophore.
o It makes the light glow green under UV light (bioluminescent).
o It is used to measure gene expression.
GUS Assay (β- glucuronidase)
o Most widely used reporter gene in plant transformation.
o GUS gene is analyzed by immersing the tissues in solution containing the substrate
X-gluc (5-bromo-4-chloro-3-indolyl-β-D-glucoronic acid).
Opine synthase
o These genes are present in Ti plasmid and both octopine and nopaline are considered
as reporters.
6. Types of Screenable Markers
Luciferase
o Fire fly based luciferase gene, a non-toxic reporter.
o The samples placed in luciferin solution in presence of ATP, oxidation emits
light captured in X-ray films.
Blue/ white screening
o It is efficiently used in identification of recombinant bacteria.
o The lacZ gene makes cells turn blue in X-gal media which is based on the
activity of β- galactosidase activity.
Chloramphenicol Acetyl Transferase (CAT)
o First reported bacterial reporter gene.
o This method is very sensitive and requires radioactive assay with substrate
labeling.
7. Selectable Markers
Selectable markers will protect the organisms against a selective
agent which kills or prevents its growth.
Since the frequency of transformation is less, the use of these
selectable marker enables researchers to make quick decisions.
The most commonly used selective agents are the antibiotics
selection.
Chloroplast killing (Eg. Streptomycin, neomycin) and
mitochondria killing (Eg. Penicillin) antibiotics are available.
8. Types of Selectable Markers
They different categories of selectable marker genes are:
o Antibiotic resistance gene markers- bacterial origin mostly
from E.coli
i. Neomycin phosphotransferase- NPT II is the most widely
used marker in plant transformation. It provides resistance to
kanamycin.
ii. Hygromycin phosphotransferase- hpt gene is used as marker
in plant transformation and more powerful than kanamycin. It
kills non-transformed cells by blocking protein synthesis.
9. Types of Selectable Markers
o Herbicide resistance gene markers
i. Bar gene- is cloned from Streptomyces hygroscopicus. Bialaphos
(secondary metabolite) contains phosphinothricin, which prevents
glutamine synthetase thereby preventing nitrogen metabolism.
Accumulation of excess ammonium in plants will result in disruption
of primary metabolism. The bar gene has modifying enzyme called
phosphinothricin acetyl transferase to prevent phosphinothricin
activity.
ii. Bromoxylin nitrilase (bxn)- it can alter the structure of herbicide
bromoxynil thereby preventing the damage to photosynthetic
machinery. It can be derived from Klebsiella ozaenue.
10. References
Selectable and Screenable Markers for Rice Transformation. In:
Jackson J.F., Linskens H.F. (eds) Testing for Genetic
Manipulation in Plants. Molecular Methods of Plant Analysis,
https://doi.org/10.1007/978-3-662-04904-4_1
https://www.biologydiscussion.com/genetics/types-of-marker-
genes-genetics/71872
https://medicine.jrank.org/pages/2491/Marker-Systems
Modes of Action of Microbially-Produced Phytotoxins,
https://doi.org/10.3390%2Ftoxins3081038
