Vectors are DNA molecules that can carry foreign DNA fragments into host cells. There are two main types of vectors: cloning vectors for propagating DNA inserts and expression vectors for expressing inserted DNA. Common vector types include plasmids, bacteriophages, cosmids, and artificial chromosomes. Vectors must have key characteristics like the ability to self-replicate, selectable markers, and origins of replication to successfully clone and transfer DNA.
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
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
vector -small fragment of DNA ,self replication,carrier or vechicle of gene of interest .vector preparation by restriction enzyme .cloning of vector by plasmids or phages .
Gene Cloning Vectors - Plasmids, Bacteriophages and Phagemids.Ambika Prajapati
A cloning vector is a small piece of DNA that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes. The cloning vector may be DNA taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium.
They allow the exogenous DNA to be inserted, stored, and manipulated mainly at DNA level.
Types -
1.Plasmid vectors.
2.Bacteriophage vectors .
3.Phagemids.
Artificial chromosome I Bacterial Artificial Chromosome I Yeast Artificial C...DevikaPatel12
https://amzn.to/3Fzcqc8
https://amzn.to/3V0q6Cp
https://amzn.to/3jbtrS7
https://amzn.to/3I0A8kr
https://amzn.to/3hrJ5s8
https://amzn.to/3FZiYlu
what is genetic engineering l genetic engineering explained l genetic engineering and biotechnology l Genetic engineeringl lbacterial artificial chromosome l p1 derived artificial chromosome l yeast artificial chromosome l Artificial chromosome l bacterial artificial chromosome vector l bacterial artificial chromosome vectors l bacterial artificial chromosomes l Bacterial artificial chromosome l yeast artificial chromosomes l yac l Yeast artificial chromosome
#BAC #YAC #bacterialartificialchromosome #yeastartificialchromosome #artificialchromosome #genetics #geneticengineering #DNA #dna #gene #genetherapy
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
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.
(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.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
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.
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.
3. VECTORS
Small DNA molecule capable of self replication.
Cloning vehicle or Cloning DNA.
Carrier of DNA fragment.
E.g. Plasmid, Phage, Hybrid vector, Artificial
Chromosomes.
5. TYPES
Two types :-
1) Cloning Vectors
Propagation or cloning of DNA insert inside a suitable host cells.
Examples: Plasmids, Phage or Virus
Obtaining millions of copies.
Uses :-
Genomic library.
Preparing probes.
Genetic Engineering Experiments.
Selection of cloning vector depends on :-
(a) Objective of cloning experiment
(b) Ease of working.
(c) Knowledge existing about the vector.
(d) Suitability.
(e) Reliability.
6. 2) Expression Vectors
Express the DNA insert producing specific protein.
They have prokaryotic promoter.
Ribosome binding site.
Origin of replication.
Antibiotic resistance gene.
Expression vectors with strong promoters.
Inducible Expression Vectors.
Eukaryotic expression vectors.
7. VECTOR
• Plasmid
• Bacteriophages
• Cosmid
• Yeast Cloning Vectors
• Ti & Ri Plasmids
TARGET HOST CELL
Bacteria, Streptomyces
Bacteria
Bacteria
Yeasts
Transformation of
cloned gene in higher
plants.
8. AGENTS USED AS VECTORS
PLASMIDS
BACTERIOPHAGES
COSMID
ARTIFICIAL CHROMOSOME VECTORS
In 1973, Cohen described first successful
construction of recombinant vector.
Plasmid PSC101 - Ecoli
9. PLASMID
Extra chromosomal DNA molecules.
Self replicating.
Double stranded.
Short sequence of DNA.
Circular DNA molecules.
Found in prokaryotes.
CHARACTERISTICS
a. Minimum amount of DNA.
b. Two suitable markers for identification .
c. Single restriction site.
d. More restriction enzyme.
e. Size range 1kg – 200kg.
f. Relaxed replication control.
g. Restriction endonuclease enzyme.
10. THREE TYPES OF PLASMID
1. Fertility plasmids:- can perform conjugation.
2. Resistance plasmids:- contain genes that build a
resistance against antibiotics or poisons.
3. Col plasmids:- contain genes that code for proteins
that can kill bacteria.
12. PBR322
Cloning Vector.
15 copies.
Reconstructed plasmid.
Derived from Ecoli plasmid- ColE1.
PBR322 – 4362 base pairs
P – denotes Plasmid
B – Scientific Boliver
R - Rodriguez
322 – number given to distinguish.
Ampilicin resistance gene derived from RSF2124.
Tetracycline resistance gene from PSC101.
Origin of replication from PMB1.
Two selectable markers
ampr tetr
14. PBR327
Derived from PBR322.
30 – 40 copies.
Expression plasmid.
Ampilicin resistance gene.
Tetracycline resistance gene.
Deletion of nucleotide between 1427 & 2516.
15. pUC8
Popular Ecoli cloning vector.
Derivative of pBR322.
Two parts derived:-
Ampicillin resistance gene.
ColEI – origin of replication.
2700 base pairs.
lac Z gene derived from Ecoli.
Polylinker sequence having unique restriction
sites
lies in lac region.
16. PHAGE
Cloning large DNA fragmance.
Linear Phage molecule.
Efficient than plasmid.
Used in storage of recombinant DNA.
Commonly used Ecoli phages :-
λ phage
M13 Phage
17. BACTERIOPHAGE VECTORS
Cloning Vectors.
It infects bacteria.
Commonly used Ecoli phages :-
λ phage
M13 Phage
Lambda phage vector
Genome size is 48,502 bp.
High transformation efficiency.
1000 times more efficient than the plasmid vector.
Origin of replication.
Genome linear in head.
Single- stranded protruding cohesive ends of 12 bases.
Cos site – site of cleavage of phage DNA.
18. Phage M13 Vectors
Filamentous bacteriophage of Ecoli.
Used for obtaining single stranded copies.
DNA sequencing.
Single stranded.
Inside host cell become double stranded.
19. HYBRID VECTOR
Component from both plasmid & phage chromosomes.
Helper phage provided.
Developed in 1978 by Barbara Hohn & John Collins.
30 – 40 Kb
Origin of replication, cloning site, marker gene, DNA cos
site.
Smaller than plasmid.
Use – construction of genomic libraries of eukaryotes.
e.g. Cosmid
20. COSMIDS
Combine parts of the lambda chromosome with parts of
plasmids.
Contain the cos sites of λ and plasmid origin of
replication.
Behave both as plasmids and as phages.
Cosmids can carry up to 50 kb of inserted DNA.
Structure of Cosmid
Origin of replication (ori).
Restriction sites for cleavage and insertion of foreign
DNA.
Selectable marker from plasmid.
A cos site - a sequence yield cohesive end (12 bases).
Ampicillin resistance gene (amp).
21.
22. ARTIFICIAL CHROMOSOME
Linear or Circular.
1 0r 2 copies per cell.
Different types –
Bacterial Artificial Chromosome (BAC)
Yeast Artificial Chromosome (YAC)
P1 derived artificial chromosome (PAC)
Mammalian Artificial Chromosome (MAC)
Human Artificial Chromosome. (HAC)
YAC – Cloning in yeast
BAC & PAC – Bacteria
MAC & HAC – Mammalian & Human cells.
23. BACTERIAL ARTIFICIAL CHROMOSOME
1st BAC Vector – PBAC108L.
Cloning of large regions of eukaryotic genome.
Origin of replication from bacterium Ecoli F -factor.
BAC vectors are pBACe3.6, pBeloBAC11.
Used in analysis of genomes.
Host for BAC is mutant strain.
24. YEAST ARTIFICIAL CHROMOSOME
Linear Plasmid Vector.
Clone large DNA segment ( 100 – 1400kb).
Occurring two forms:-
Circular – grows in bacteria.
Linear – multiplies in yeast cells.
pYAC3 - first YAC developed.
It contains :-
ARS sequence – replication
CEN4 sequence – centromeric function
TRP1 & URA3 – 2 selectable markers
Use – mapping complex eukaryotic chromosome .