Name : Dina Ulyana SN

Student Number : 1706103010011

Class : 04 (Pgmipau)
Course : Biotechology
Lecturer : Iswadi, S.Pd., M.Si
Date : December, 12, 2020

The First Video Resume

THE DIFFERENCE BETWEEN TRANSFORMATION, TRANSDUCTION, AND

CONJUGATION

The transformation process is a process in which DNA or DNA fragments are taken
directly by adjacent bacteria. In transduction of all DNA, generally the genetic material is DNA
so that DNA is transferred by bacteriophage viruses from one bacterium to another. What is the
conjugation of this one and the third is when genetic transfer occurs through direct contact
between bacteria. These are three different situations that are known, now in all these types of
gene transfer there are two types of organisms involved with one donor (giver), another recipient
(donor and recipient). Donor giving, gene recipient received in all three cases.

Differences on a large/major scale:

Transformation: Collection of naked DNA by bacteria

The first major scale difference is the optical transformation which is a bare DNA
fragment (DNA without cells or associated proteins) by bacteria. For example, usually what
happens when the donor cell starts to die, DNA fragmentation begins to occur in the bacterial
cell and said job apoptosis or cell death all those DNA fragments that enrich the bacterial
chromosomal DNA begin to degrade. As a cell, the cell membrane wall breaks away from the
bacteria so that immediately the environment is filled with DNA fragments that come from the
donor cell. In the same environment there are cells receiving the same cells from the same
progeny cells from the same generation. Recipient cells can sense an environment filled with so
many fragments of DNA content from the DNA that they can easily pick up any of the fragments
inside the cell. After this retrieval, the fragment can be recombined with the bacterial
chromosome of the recipient cell. If it is recombined then the new gene sequence involved and
inserted into the recipient cell with the help of this new DNA which is combined here they can
have different features. The next one is transduction.
Transduction: Bacterial DNA transferred by viruses (phages)

Transduction is a process in which DNA is transferred from one bacterium to the next
with the help of a virus. It also refers to the process by which foreign DNA is introduced into
other cells via viral vectors. So this is the scenario when the iris pouch infects the bacteria it
injects its DNA. Inside the bacteria and then the virus starts to break down the bacterial
chromosomes into fragments. Now fragmentation takes place in donor (giver) cells. Now that
they have started packing the genetic material inside the head of the virus or particles of the viral
head sometimes by mistake, they take some part of the bacterial chromosome or bacterial DNA
structure in one of these viral heads. Now the filler head carrying the breakdown content of the
bacterial DNA can be transferred because the fudge can go ahead and sit down and inject its
DNA into the recipient cell. The recipient cell will receive a copy of the other bacterial
chromosome for the bacterial DNA. Now, that DNA can be recombined with the recipient cells
so they can have this way of essentially transforming without the virus from bonding with the
virus and the process is similar to the only transfer of bacterial DNA fragments but in their
transduction using viruses. To do that in transformation, they took it right away.

Conjugation: Transfer of DNA between bacterial cells

But the third one is a very different conjugation which is a kind of direct interaction
between two bacteria and adjacent bacteria. They engage with each other with the help of a
tunnel that forms between them which is known as the pilus or f.pilus is a conjugation cube that
is formed between the two adjacent cells and through this tube they can transfer their genetic
content they can transfer it in the plasmid of one bacterium. to other bacteria. Conjugation
process, having one donor, and having one recipient. The donor has F pilus. F pilus means F
stands for fertility here and F plasmid which is called F pilus needs fertility plasmids and bacteria
containing F plasmids are always donors because they will donate this F plasmid to the recipient
cells. So, those who carry this plasmid, in this video, are called male types. It resembles the male
and ISM recipient to the female in this respect and the whole process resembles that of sexual
reproduction but not exactly. In this case with the help of the spiller it is not attached to the
receiver and they form a tube in the middle and this tube is known as conjugation. They form the
conjugate tube in the middle after forming the tube then they can easily start to replicate this
plasmid F with the help of this rolling loop mechanism with the help of this rolling loop
replication they start the replication and meanwhile they transfer this Plasmid content through
the conjugation tube to the adjacent bacterial cell and then after this whole process is complete.
Both cells will receive one copy of the plasmid if each is due to a plug that was previously
present in the donor but now the recipient will get one plasm. mid so after getting the plasmid it
gave the characteristics of the simplest make and now can re-interact with multiple recipients and
do the same thing. It is a general description of conjugation when in trance formation has DNA
fragments produced after the death of a bacterium.
 Once the bacteria die, cellular components are destroyed and DNA fragments float in the
environment. So other bacteria nearby come in it save the environment that there are fragments
on the outside so there's a chance some of these fragments might contain any kind of antibiotic
resistant gene. If it carries antibiotic resistance and somehow this recipient can pick up a DNA
fragment containing a gene that is resistant to antibiotics then this recipient bacterium can
acquire antibiotic resistance too, that's the idea of transfromation. In this case they will take the
DNA fragment and will join the recipient cell so after taking this fragment they enter the DNA
fragment into the cell with the help of recombination. Its use is a homologous recombination
process to properly place this DNA in its genome with these types of types they converge on
many different features. The characteristics of producing different enzymes such as enzymes that
produce various types of chemicals or enzymes that will fight antibiotics and also antibiotic
resistance factors. . Then the DNA of the host bacteria is degraded by the charge so that it is so
satisfied to do so after that the DNA quickly begins to replicate producing further genetic
material. Once they produce so much genetic material rapidly then they process that material and
put it into a new hereditary mode capsid during this packaging which is known as packaging.
DNA packaging is deep in the inner head, during this packaging because of some mistakes and
problems sometimes they also put some of this bacterial DNA into the head of fashion. Now this
is rare but it happens as for example in the image in this video, one in five large particles receive
DNA back. Now the post-plant transport and transfer are contained within other bacteria which
are the host bacteria. It inserts into the recipient bacteria which injects the DNA in it, so now this
recipient bacteria receives the DNA content which should be fake content but in this case it is the
bacterial DNA content from the donor in this way the recipient bacteria receives the part from
the donor. This bacterial DNA means that the DNA can be fused with the genome of the
recipient bacteria, this fusion may be complete and if new gene requirements are obtained. So in
all these processes there are different processes involved but all these processes are all needed
especially for the collection and formation of antibiotic resistance properties by bacteria so that
in the sense is the difference between conjugation and transduction transformation.
Second Video Resume

GENE TRANSFER METHODS AND TECHNIQUES IN PLANTS

The transfer of genes from one organism to another is a natural process that creates a
variety of biological traits. This fact underlies all efforts to increase agro-important species,
either through traditional agricultural breeding or through molecular biology techniques. In both
cases, humans manipulate naturally occurring processes to produce varieties of organisms that
exhibit desired traits, for example, animal feed with a higher proportion of muscle to fat, or
disease-resistant corn. Laboratory methods for transferring individual genes between organisms
take advantage of naturally occurring gene transfer mechanisms in addition to sexual
reproduction. This includes the uptake of DNA by cells and cell-to-cell transfer of packaged
genetic material such as viruses.
Many factors must be considered in the design of a gene transfer system. The first
requirement is an easily detectable "tag" for the gene so that its development into a new host can
be traced. Another important consideration is the efficiency of gene transfer. The probability of
success must be high enough for gene transfer to be detected with a reasonable frequency. If
drug resistance or other selection schemes are used, lower frequencies may be acceptable.
Special vectors can increase the efficiency of gene transfer. Foreign genes attached to the vector
will be carried by them into the host cell. Vectors often come from coiled DNA molecules called
plasmids, or from viruses.

Several methods are used for gene transfer in plants

1. Direct DNA Collection
DNA is inserted into plant cells that are cultivated in the laboratory is the direct extraction of
DNA from the surrounding culture media. DNA must enter the cell and be maintained stably
and be inherited in the cell line in such a way that its new genetic information is expressed to
give the cell new traits. The main advantage of direct DNA retrieval is its simplicity and
application to many organisms and cell types.

2. Chemical Treatment
The cells are mixed with DNA which has been precipitated with calcium phosphate. This
treatment compresses the DNA, so that the cell takes multiple copies of the foreign gene.
Alternatively, the chemical DEAE-dextran could be used to facilitate DNA retrieval.
Polyethylene glycol has been used to obtain direct uptake and stable maintenance of DNA
by protoplasts of wheat species, Triticum monococcum, other monocot grasses, Lolium
multiforum.

3. Electroporation
The cell is mixed with the DNA in solution and is subjected to a short electric current.
Electroporation can work for all types of cells, even those that have resisted taking DNA by
 chemical treatments, for example, immune system cells. Electroporation can insert DNA
into protoplasts of the two main categories of plants - dicots (eg carrots and tobacco) and
monocots (eg corn). Electroporation provides a temporary gene expression system for
plants. Electroporation also allows stable integration of genes into plant chromosomes. It has
been used successfully to stably transform maize and tobacco cells.

4. DNA Micro Injection

Micro injection can be used to deliver genetic material into plant cells. DNA segments,
entire chromosomes, and even cellular organelles such as chloroplasts, which contain their
own DNA molecule, can be micro-injected by the methods used for animal cells, although
certain physical properties of plant cells complicate this technique. Microinjection of
individual chromosomes or cellular organelles (for example, chloroplasts, mitochondria, and
nuclei) has the potential to produce better cultivars with novel traits such as herbicide
resistance or cytoplasmic male sterility. Trait transfer by microinjection is more direct,
precise, and faster than by cell culture or fusion, because microinjection transfers specific
and limited genetic information. There will be little need for selection or backcrossing,
which is often a time consuming, arduous process. The micro-injection of chromosomes will
also allow the transfer of traits encoded by a single gene that has not been identified and
isolated.

5. Cell Fusion
Cell fusion combines all of the genetic contents of the two cells, producing hybrid cells that
often express certain traits from both parents. Stem cells can come from different species or
from different types of the same species. Fusion is usually mediated by chemicals such as
polyethylene glycol or dimethylsulfoxide, although newer techniques use electrofusion. In
eukaryotic cells, the cytoplasm of the part of the cell that surrounds the nucleus contains
organelles that have separate DNA. In plants, protoplast fusion is used to transfer genes from
the nucleus and cytoplasm. Fusion combines the genomes of two hosts, as in traditional
breeding, but yields can sometimes be obtained more quickly, although fusion products must
be crossed back into the receiving line over several generations to create a new, stable line
that has one desired trait of the donor. Protoplast fusion can be used to transfer genes that are
difficult to identify, isolate, and clone or for polygenic traits. Furthermore, protoplast fusion
can be used for plants that cannot be crossed sexually (although plants that regenerate from
fused hybrids are sometimes sterile).

6. Vector-Mediated Gene Transfer

Vector is a DNA molecule that binds to a foreign gene to facilitate its transfer, maintenance,
and expression within the target cell. Vectors offer the following advantages: high frequency
of gene transfer, transfer to specific cell types, more control over the number of final copies
of the transferred genes, and certain traits that make them traceable, enabling them to be
maintained stably in target cells, and enabling them to express foreign genes. Therefore,
 vectors can greatly enhance gene transfer. However, different species and cell types may
require different types of vectors, and often a lot of work must be done to create a suitable
vector system before genes can be transferred to a particular organism.
a. Cauliflower Mosaic Virus
small double strand DNA viruses that infect cruciferous plants, such as cabbage and
mustard greens. CaMV is transmitted in nature by aphids, but its DNA can infect plants
if only applied to their leaves. CaMV causes systemic infection and replicates in
abundance throughout the plant.

b. Gemini Virus
Gemini virus is a plant single-stranded DNA virus that is transmitted by insects, such as
leafhoppers. Viruses in this group infect many crops, including wheat and corn monocots
as well as dicot beans, tobacco, and tomatoes.

c. RNA viruses
DNA cannot be carried on RNA directly. However, scientists can make complementary
DNA copies of the RNA viral genome. This copy can be used to create vectors that will
carry foreign genes. The DNA can then be transcribed back into RNA, allowing the
engineered virus to infect cells. There are several potential advantages of RNA virus
vector systems for plants. First, once infected with cloned DNA or RNA transcripts in
vitro, plants must immediately express these new properties, in contrast to the Ti plasmid
system (discussed in the next section), where a long regeneration process is usually
required to achieve the transformation of the plant. Second, the expression of viruses as
self-replicating extrachromosomal RNA molecules means that gene expression will not
be affected by a `` position effect '' due to insertion at an undesirable site in the plant
chromosome. Third, gene expression via a powerful viral promoter, combined with
template replication to produce multiple copies of the gene, will enable the production of
a large number of specific gene products in plant cells.

d. Transposable elements
Transposable elements (also called "transposons") can move from one place to another
in an organism's genome and pick up extra pieces of DNA to carry. These elements share
some of the same physical and functional properties as retroviruses, but they do not
spread from cell to cell via infection and are therefore not considered viruses.
Transposons have been used to modify the Pseudomonas fluorescens bacteria that live in
corn roots by inserting the insecticide gene from Bacillus thuringiensis.

e. Ti Plasmid
 Plasmids are circular DNA molecules that exist independently of the main chromosome
of the cell; Ti plasmids are a fairly large natural variety. Agrobacterium infects most
dicot species and causes a tumor disease called crown gall. The disease is triggered by
the transfer of natural genes from the bacterial Ti plasmid portion, called T-DNA, into
the plant chromosomes. Plants that can currently accept the Ti plasmid vector include
petunias, tobacco, soybeans, carrots, tomatoes, alfalfa, and oilseed rapeseed. The
transferred gene includes a small subunit of the plant photosynthetic enzyme ribulose
1,5-bisphosphate carboxylase.

f. Fungal and Bacterial Plasmids

Fungi and pathogenic bacteria can be used as biological control agents for insect pests or
weeds. The isolation and transfer of pathogenicity genes serves two purposes: the
development of enhanced agents for biological control, and the discovery of resistance
genes in plants or insects that fight pathogenicity. The use of transposons and plasmids
to isolate and study pathogenic genes from fungi and bacteria that attack plants will lead
to understanding the molecular basis of many agronomically critical diseases and
suggesting ways to combat them.
The Third Video Resume

GENE TRANSFER TECHNIQUES IN PLANTS AND ANIMALS

The newly inserted DNA sequences that lead to the formation of transgenic organisms
are called trans genes, for successful integration and expression, the transgene must have certain
proponents, they are the promoter safeguards for the correct expression of the genes transferred
of interest. The genes that you really want to insert are the ones called transgenes.

GMO Crops
Transgenic plants that contain a single gene or group of genes that have been specifically
introduced into plants instead of plants. Getting it through the natural process of pollination, the
trans gene that is inserted can come from various sources, such as other unrelated plants or from
completely unrelated species such as bacteria or animals or even humans. Methods of
transferring genes into plants There are several methods of transferring genes into plant cells,
broadly divided into three categories, namely physical methods, chemical methods, and
biological methods. Physical methods and chemical methods of gene transfer are also
collectively referred to as vectorless gene transfer or direct gene transfer. The biological method
of gene transfer is vector-mediated gene transfer or indirect gene transversion. In this method,
the desired gene is carried to the target cell by a molecular vehicle called a vector, so that we
have a physical, chemical and biological method of transferring genes into plants, let's consider
one by one starting with the physical method. The main physical methods used in plant
transgenesis include particle gun, electroporation, and micro injection. This method of gene
transfer is referred to as biological gene transfer. in this method the foreign DNA containing the
gene to be transferred is coated onto the surface of very small gold or tungsten particles. As these
particles are only one to three micrometers in size, they are then bombarded into target tissues or
cells using particle weapons.

Electroporation
Electroporation as the name implies involves the generation of pores using an electric
current, when a high-voltage electric pulse is applied to the plant cell, transient pores or holes are
formed on the surface of the plant cell. When a plant cell is placed in a medium containing DNA
for transport, the DNA enters the cell through these pores into the cytoplasm and further into the
nucleus. thus we have succeeded in moving the DNA from the medium into the cell. Generally,
for gene insertion using protoplast electroporation, protoplasts are none other than plant cells that
do not have a hard cell wall. Although protoplasts are preferred to use, this technique is also
applied to all plant cells and plant tissues.
In the next technique which is micro injection in this technique the DNA is pulled into a
very special nano syringe or even this smaller femtosyringe which is done with the aid of a
microscope, in fact the whole procedure is done looking through a high place. The microscope
definition of the cell to be converted is held in position using a nanoscale suction pipette and
DNA is carefully injected into the cells. In this way we can ensure that the DNA actually reaches
the cell, in this technique the DNA insertion can also be made without a marker gene that can be
selected so as to produce a marker free transgenic plant.

Gene Transfer Chemical Methods

Some chemicals can cause cell membrane instability, resulting in uptake of the desired
DNA molecules. Commonly used chemicals including polyethylene glycol and dextran sulfate,
calcium phosphate, have also been used in several studies, but are the main chemical methods
used to use liposomes. Liposomes or liposomes as they are also called are structures built with
lipid molecules, spherical in shape with an aqueous nucleus, if the liposomes are built up in an
aqueous medium containing the genes to be transferred. Some of the genes are trapped inside the
liposomes, these DNA-loaded liposomes can then be added to the plant cell where they fuse with
the membrane and release their contents or desired genes into the cell. for this technique also
protoplasts are preferentially used.

A Biological Method of Transferring Genes Into Plants

The biological method of gene transfer is also referred to as the vector-mediated gene
transfer method. The biological method most commonly used in plants is agrobacterial
intermediate gene transfer. agrobacterium tumefaciens are soil bacteria that cause tumor-like
growths called crown gall on plants. this is the result of the transfer of the plasmid from bacteria
and this plasmid is called the tumor-inducing plasmid tior.

Physical Methods of Transferring Genes To Animals

The main method of transferring genes into animals by physical means is electroporation.
the principles of this method are the same as those used for plants. because animal cells do not
have a strong cell wall, the need for the manufacture of protoplasts or other similar structures
does not arise so this method is preferred for those used with animal cells. The micro injection
technique has been used successfully to transfer trans genes into animal cells. However, more
often the technique of micro injection in animal cells has been used to transfer the entire pro
nucleus rather than a small piece of DNA, in this method the fertilized zygote is the e-nucleus i.e.
the nucleus is removed from the zygote by suction and instead of the nucleus.

Chemical Methods of Transferring Genes To Animals

Chemical methods of transferring genes into animal cells of polyethylene glycol or
calcium phosphate can be used to shake the membrane for DNA retrieval. The transfer method is
the same as the method for plant protoplasts, but the more commonly used chemical method for
transferring genes into animal cells, is the spread of liposomes that are laden with the desired
gene because they readily fuse with the animal cell membrane and release them. DNA into cells.
several vector-mediated gene transfer methods have also been devised for animal cell
transgenesis. Among the most important are retroviral transfer, transgenesis by engineering
embryonic stem cells, and transgenesis with artificial high-capacity vectors.
Retroviral Transfer
Retrovirus particles which are highly contagious are transformed into a state of disrepair.
this modification deprives them of their infective or pathogenic properties. however they retain
their capacity to transfer DNA into animal cells. helper virus strains that can carry large chunks
of inserted DNA can also be used to transfer genes.

ES Cell Engineering For Gene Transfer

Using embryonic stem cells during the development of an animal fetus in the blastocyst
stage the cells can be harvested and maintained in culture. these cells can reproduce very well
under cultured conditions and retain their ability to differentiate into various cell types. for this
reason they are called pluripotent stem cells or simply embryonic stem cells or ES cells.

GMOs With Artificial High Capacity Vectors

Although some of the methods discussed so far are very effective at transferring short
segments of DNA, they are inadequate for transferring the large chunks of DNA that are often
desired in animal transgenesis. because these high-capacity vectors are constructed in an
artificial way, these are collectively called artificial chromosomes. The main artificial
chromosomes include dorsal or bacterial artificial chromosomes, p1 packets or bacteriophages
derived by artificial chromosomes, yeast or yeast artificial chromosomes, max or mammalian
artificial chromosomes, and man-made hacks or chromosomes. of all types of chromosomes
yeast or yeast is the most widely used vector.