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Unit 18 Assignment 4 Genetic Engineering
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Assignment 4Genetic Engineering is the process of changing the genetic makeup of an organism whether it’d be animal, plant or bacterium
through using technology. In genetic engineering an organism’s genetic material is either altered or selected in a hope that the
organism will obtain specific characteristics. Genetic engineering does have a lot of controversy against it however, it does also
serve many purposes which are rather useful. Genetic engineering was brought about during the 1960’s however, major
experiments failed to become recognized until around the 1990’s.
One type of genetic engineering is cloning. Cloning is a process of producing a genetically identical copy of an organism using
genetic material like DNA. Even though this is possible through the process of asexual reproduction whereby organisms like
bacteria and certain plants are able to create offspring which is genetically identical to the parent. To create clone’s modern
genetic technology is also able to be used by scientists. There are three types of cloning these are; gene cloning, reproductive
cloning and therapeutic cloning.
Gene Cloning - Gene cloning also known as DNA cloning is a process of cloning whereby a gene of interest becomes located and
cloned (copied) out of DNA which has been extracted from an organism. In simpler terms it is the process of making multiple,
identical copies of a certain piece of DNA. When extracting DNA from of organism all its genes will be extracted at one time. The
DNA will consist of thousands of various genes as this is the case the genetic engineer needs to find the specific gene which
encodes for the specific protein which is of interest. In a general procedure of gene cloning, the gene or other DNA fragment
which is of interest will be inserted firstly into a plasmid which is a circular piece of DNA. The DNA fragment is inserted into a
plasmid using enzymes which essentially ‘cut and paste’ the DNA producing a molecule of recombinant DNA this is DNA which is
assembled out of fragments from various sources. Here, a circular piece of plasmid DNA which has overhangs on its end are a
match to that of a DNA fragment. The DNA fragment and the plasmid become joined to one another to produce a gene-
containing plasmid, an example of recombinant DNA. The recombinant plasmid then becomes introduced into bacteria. The
bacteria which carries the plasmid is selected and grown up so, when they reproduce, they replicate the plasmid and pass this
on to their own offspring making copies of the DNA which it consists of. A fragment of DNA encodes a useful protein and the
bacteria is used as a way to make the protein. An example of this is expressed in the E. coli bacteria is the human insulin gene
which has the ability to make insulin which is used by diabetics. When cloning DNA to synthesize a protein in bacteria there are
certain steps which are usually followed as these are as follows;
Method-
Firstly, the plasmid needs to be cut open and “paste” the gene in. The primarily relies on two main things which
are restriction enzymes and DNA ligase. A restriction enzyme is a type of DNA-
cutting enzyme which has the ability to recognize a certain target sequence and
cuts DNA into two pieces either near or at the specific target sequence site. These
restriction enzymes tend to produce cut ends which have short and single-stranded
overhangs. Therefore, if two molecules have overhangs which match, they are able
to base-pair and stick together, this is where DNA ligase comes in. As without the
DNA ligase they won’t be able to combine to create an DNA molecule which is
unbroken so, when they come joined by the DNA ligase it seals the gaps present in
the DNA backbone. The aim is to insert a target gene into the plasmid. Both the
target gene and the plasmid will need to be separately digested along with the
restriction enzyme. The fragments are combined and purified and have single-
stranded DNA which overhands which gives them the ability to stick together. The
DNA ligase joins the fragments with matching ends to one another to create a
single molecule of DNA which is unbroken. Producing a recombinant plasmid what has the target gene.
Then, through a process known as transformation plasmids and other DNA is able to
be introduced into bacteria. Bacterial cells which have been specially prepared are
now given a shock e.g. high temperature which makes them more apt and
encourages them to take up the foreign DNA. Here, only a small minority of the
bacteria will take up a plasmid successfully. A plasmid generally contains a gene which
is antibiotic resistance therefore, this allows survival of bacteria when in the presence
of certain antibiotics. So, bacteria which took up the plasmid is able to be selected on
a nutrient plate which contains the antibiotic.
Without a plasmid the bacteria will die however, the
bacteria which carries a plasmid is able to live and
reproduce. All the bacterium which survives will produced a small colony of
identical bacteria which will carry the same plasmid. Concequently, not all the
colonies may contain the correct plasmid as during ligation the DNA fragments
don’t always get ‘pasted’ in the way which was intended. Due to this, DNA
needs to be collected from more than one colony as this will determine
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