Bacterial transformation involves bacteria taking up foreign DNA from their environment. This process allows bacteria to horizontally transfer genes. Transformation is used in DNA cloning to introduce plasmids containing genes of interest into bacteria. The bacteria are then selected using antibiotics to identify those containing the plasmid, which can then be used to produce large quantities of the gene or its protein product.
What are an expression vector? Detailed description of plant gene structure. Plant expression vector systems are generally consists of Ri and Ti plasmids.
The other vectors which are generally used are DNA and RNA viruses.
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.
What are an expression vector? Detailed description of plant gene structure. Plant expression vector systems are generally consists of Ri and Ti plasmids.
The other vectors which are generally used are DNA and RNA viruses.
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.
Sanger sequencing is a method of DNA sequencing based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
in this presentation, what are the steps and strategies involved the gene cloning and i was focused only on the 1st two steps of gene cloning.they are generation of foreign DNA molecules and selection of suitable vectors.
Sanger sequencing is a method of DNA sequencing based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
in this presentation, what are the steps and strategies involved the gene cloning and i was focused only on the 1st two steps of gene cloning.they are generation of foreign DNA molecules and selection of suitable vectors.
Prokaryotes can exchange DNA with eukaryotes, although the mechanisms behind this process are not well understood. Suspected mechanisms include conjugation and endocytosis, such as when a eukaryotic cell engulfs a prokaryotic cell and gathers it into a special membrane-bound vesicle for degradation.
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This will be used as part of your Personal Professional Portfolio once graded.
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Prepare a presentation or a paper using research, basic comparative analysis, data organization and application of economic information. You will make an informed assessment of an economic climate outside of the United States to accomplish an entertainment industry objective.
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The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
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http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
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1. NAME – SAMADRITA BANIK
ST. GEORGE COLLEGE OF MANAGEMENT
AND SCIENCE
M.Sc MICROBIOLOGY
2nd SEMESTER
2.
Bacterial transformation is a process of horizontal gene
transfer by which some bacteria take up foreign genetic
material (naked DNA) from the environment. It was first
reported in Streptococcus pneumoniae by Griffith in 1928.
DNA as the transforming principle was demonstrated by
Avery et al in 1944.
INTRODUCTION
3.
Bacteria can take up foreign DNA in a process called
transformation.
Transformation is a key step in DNA cloning. It occurs after
restriction digest and ligation and transfers newly made
plasmids to bacteria.
After transformation, bacteria are selected on antibiotic plates.
Bacteria with a plasmid are antibiotic-resistant, and each one
will form a colony.
Colonies with the right plasmid can be grown to make large
cultures of identical bacteria, which are used to produce
plasmid or make protein.
BACTERIAL
TRANSFORMATION
4.
Transformation and selection of bacteria are key steps in DNA cloning. DNA cloning is the
process of making many copies of a specific piece of DNA, such as a gene.
The copies are often made in bacteria.In a typical cloning experiment, researchers first insert
a piece of DNA, such as a gene, into a circular piece of DNA called a plasmid.
This step uses restriction enzymes and DNA ligase and is called a ligation.After a ligation,
the next step is to transfer the DNA into bacteria in a process called transformation.
Then, we can use antibiotic selection and DNA analysis methods to identify bacteria that
contain the plasmid we’re looking for.
DNA CLONING
6.
Specially prepared bacteria are mixed with DNA (e.g.,
from a ligation).
The bacteria are given a heat shock, which causes some of
them to take up a plasmid.
Plasmids used in cloning contain an antibiotic resistance
gene. Thus, all of the bacteria are placed on an antibiotic
plate to select for ones that took up a plasmid.
8. Bacteria without a plasmid die. Each bacterium with a plasmid
gives rise to a cluster of identical, plasmid-containing bacteria
called a colony.
Several colonies are checked to identify one with the right
plasmid (e.g., by PCR or restriction digest).
A colony containing the right plasmid is grown in bulk and
used for plasmid or protein production.
9.
Possibility 1: Bacteria = plasmid factories
In some cases, bacteria are simply used as "plasmid factories," making lots of
plasmid DNA. The plasmid DNA might be used in further DNA cloning steps
(e.g., to build more complex plasmids) or in various types of experiments.
In some cases, plasmids are directly used for practical purposes. For instance,
plasmids were used to deliver a human gene to lung tissue in a recent gene
therapy clinical trial for patients with the genetic disorder cystic fibrosis
Protein production in bacteria
10.
Possibility 2: Bacteria = protein factories
In other cases, bacteria may be used as protein factories. If a plasmid contains
the right control sequences, bacteria can be induced to express the gene it
contains when a chemical signal is added. Expression of the gene leads to
production of mRNA, which is translated into protein. The bacteria can then be
lysed (split open) to release the protein.
11.
Bacteria contain many proteins and macromolecules. Because of
this, the newly made protein needs to be purified before it can be
used. There are a variety of different techniques used for protein
purification.
12. In one technique called affinity chromatography, a mixture of molecules
extracted from the lysed bacteria is poured through a column, or a cylinder
packed with beads. The beads are coated with an antibody, an immune
system protein that binds specifically to a target molecule.
The antibody in the column is designed to bind to our protein of interest, and
not to any other molecules in the mixture. Thus, the protein of interest is
trapped in the column, while the other molecules are washed away. In the
final step, the protein of interest is released from the column and collected
for use.
13.
Transformation is the process by which foreign DNA is introduced
into a cell. Transformation of bacteria with plasmids is important
not only for studies in bacteria but also because bacteria are used as
the means for both storing and replicating plasmids. Because of
this, nearly all plasmids (even those designed for mammalian cell
expression) carry both a bacterial origin of replication and an
antibiotic resistance gene for use as a selectable marker in bacteria.
CONCLUSION