1. Yeast plasmids like the 2 micron circle have been extensively studied and developed into yeast cloning vectors.
2. Shuttle vectors like YEp vectors contain selectable marker genes like LEU2 and bacterial plasmid origins of replication like pBR322, allowing them to replicate in both E. coli and yeast.
3. The 2 micron circle is a 6kb endogenous yeast plasmid that replicates autonomously through an ARS sequence and is maintained at 50-100 copies per cell.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
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
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.
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
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.
gene cloning in eukaryotes (gene transfer).pdfNetHelix
Gene cloning recently faced difficulties associated with
bacteria, especially when dealing with
genes from eukaryotic organisms so we should to employ the eukaryotic expression in this PDF we will learn about gene cloning in eukaryotes, types of yeast plasmids and the importance of each one
systems
A genetically engineered DNA molecule from bacteria , phage or yeast to carry foreign DNA for the purpose of cloning and expression of the inserted DNA of interest in RDT
Fungal Transformation in yeast and filamentous fungi
Introduction to Fungi
Background of fungal transformation
Transformation protocol
Transformation vectors
Integration into chromosomes
Biological applications of fungi
Conclusion
References
• Plasmids are extra-chromosomal genetic elements that replicate independently of the host chromosome.
• They are small, circular (some are linear), double-stranded DNA molecules that exist in bacterial cells and in some eukaryotes.
“The technique in which a DNA fragment carrying a gene inserted into the cloning vector, this vector is introduced in the living cell and subsequent propagation of this recombination DNA molecule into many copies is known as gene cloning.
description of plasmids and types and importance of plasmids and artificial plasmids(PBR322,cosmids,phagemids) and selection of the recombinants and uses and advantages and disadvantages of the plasmids
Transgenic animal production and its applicationkishoreGupta17
A genetically modified animal with the heterologous gene of interest being inserted for the purpose of biopharming or make a diseased model to study the consequences of disease and its probable therapy
for cloning and expression of exogenous gene or gene throthrough vector it must be introduced into the host cell through transformation , ,transduction, electroporation gene gun etc.
A cytological technique to detect the nature of adjacent chromosomal regions by using different staining technique assisted with some pre treatment of metaphase chromosomes prepared on the slides
How to Make a Field invisible in Odoo 17Celine George
It is possible to hide or invisible some fields in odoo. Commonly using “invisible” attribute in the field definition to invisible the fields. This slide will show how to make a field invisible in odoo 17.
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
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A Strategic Approach: GenAI in EducationPeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
Honest Reviews of Tim Han LMA Course Program.pptxtimhan337
Personal development courses are widely available today, with each one promising life-changing outcomes. Tim Han’s Life Mastery Achievers (LMA) Course has drawn a lot of interest. In addition to offering my frank assessment of Success Insider’s LMA Course, this piece examines the course’s effects via a variety of Tim Han LMA course reviews and Success Insider comments.
Acetabularia Information For Class 9 .docxvaibhavrinwa19
Acetabularia acetabulum is a single-celled green alga that in its vegetative state is morphologically differentiated into a basal rhizoid and an axially elongated stalk, which bears whorls of branching hairs. The single diploid nucleus resides in the rhizoid.
2. Yeast Plasmid
• Plasmids are not limited to bacteria.
• For example, some plasmids have been
extensively studied in yeast and developed
into yeast cloning vectors.
• These plasmids have also been used as
"simple systems" to understand the
mechanism and control of DNA replication in
eukaryotic cells.
3. Introduction
• A shuttle vector is a vector (usually a plasmid) constructed
so that it can propagate in two different host species.
• DNA inserted into a shuttle vector can be tested or
manipulated in two different cell types.
• Also called bifunctional vector since in two host
5. Features of yeast plasmid
• Yeast has genome about 2X10 7 bp constituting 16 linear
chromosomes,
• Some strains posses a type of plasmid known as the 2 micron circle
containg 6318 bp and in number 70-200 copies
• Yeast hasfour groups depending on replication mode
• YIp- Replicate by integration since lack independent origin for
replication . Yip –one copy /cell – stable , segment of pBR322with ampicilin
+ and bacterial origin
• YRp- Yeast replicating plasmid – has indepent ARS ( autonomus
replicating sequence , but unstable
• YCp- yeast centromeric plasmid- has centromeric sequence that
can attach spindle fibres ( like chromosomes )
• YEp-Yeast episomal plasmids (YEps) are shuttle vectors. They can
replicate in E. coli and also in yeast. Yep can replicate as an
independent plasmid, but also implies that integration into one of
the yeast chromosomes can occur. Number 20-200 /cell
• ... all can be maintained in both bacteria & yeast
6. 2 micron plasmid
• This plasmid is found in several strains of yeast,
Saccharomyces cerevisiae.
• It is 6 kb in size and has a copy number between 70 and 200.
• It replicates autonomously by using several enzymes provided
by the host cell and the proteins coded by its own genes.
• The 2 Micron (µm) Plasmid, 2. LEU2 Gene, and 3. The pBR
322.
• The 2µm plasmid has an origin of replication and two genes
involved in replication, and also encodes a site-specific
recombination protein, called FLP, which is homologous to the
phage λ, integrase and can invert part of the 2µm DNA
sequence
7. 2u circle
• One interesting yeast plasmid is called the 2u circle. The 2u
circle is a 6.3 kb circular, extrachromosomal element found
in the nucleus of most Saccharomyces cerevisiae strains.
• The 2u circle doesn't give cells that carry it any apparent
selective advantage, but it is stably maintained at about 50
to 100 copies per haploid genome of the yeast cells.
• Like the host chromosomes, the 2u circle is coated with
nucleosomes and replication is initiated by host replication
enzymes once per cell cycle.
• The origin of bidirectional DNA replication is initiated at a
specific site on the plasmid called an ARS sequence
("autonomous replication sequence").
8. ARS
• An autonomously replicating sequence (ARS) contains
the origin of replication in the yeast genome. It
contains four regions (A, B1, B2, and B3), named in
order of their effect on plasmid stability. The A-Domain
is highly conserved, any mutation abolishes origin
function.
• Mutations on B1, B2, and B3 will diminish, but not
prevent functioning of the origin.
• Element A is highly conserved, consisting of the
consensus sequence:
• 5'- T/A T T T A Y R T T T T/A -3‘
• Y is either pyrimidine and R is either purine
9. LEU2 gene
• LEU2 gene is a gene of yeast chromosome. It codes for an enzyme
called isopropyl malate dehydrogenase which is involved in
conversion of pyruvic acid to leucine.
• When LEU2 gene is used as a selectable marker, the host yeast
must be a mutant with a non-functional LEU2 gene. Such strains of
yeast are called leu2.
• These mutant cells are unable to synthesize leucine and will grow
only if this amino acid is added to the growth medium.
• However, yeast cells transformed by the YEp (yeast episomal
plasmid) containing LEU2 gene can grow in the growth medium
lacking the amino acid leucine.
• Thus, the transformed yeast cells (called transformants) can be
selected.
10. . The pBR 322
It is the bacterial plasmid. It has two marker genes:
the ampicillin and tetracycline resistant genes (i.e.,
AmpR and TetR). This plasmid has its own origin of
replication.
Hence, initial gene manipulation experiments can be
done with E. coli.
The YEP vector is episomal in nature as it can integrate
with one of the yeast chromosomes. Integration
occurs by homologous recombination between the
LEU2 gene. This results in the insertion of the entire
YEP into on of the yeast chromosome.