REPORT IN

BOITECHNOLOGY
Objectives:
-Using diagram, illustrate how bacterial
plasmids are used as vectors for the transfer of
genes from one cell to another.
-Explain the role of restriction enzymes and
DNA ligase in creating recombinant DNA.
-Explain the importance of gene cloning.
-Describe how to construct a library of genes of
genes from organism.

Reporters:
Sheryl Galario
Eunice Ruda
Teacher:
Ms. Wawie Mindoro

Bacterial plasmids as vectors

Artificially constructed plasmids may be used as vectors in genetic
engineering. These plasmids serve as important tools in genetics and
biotechnology labs, where they are commonly used to clone and amplify
(make many copies of) or expressparticular genes.[11] A wide variety of
plasmids are commercially available for such uses. The gene to be replicated
is normally inserted into a plasmid that typically contains a number of
features for their use. These include a gene that confers resistance to
particular antibiotics (ampicillin is most frequently used for bacterial strains),
an origin of replication to allow the bacterial cells to replicate the plasmid
DNA, and a suitable site for cloning.

What role does DNA ligase play during replication?

Ligase is an enzyme which fills the gap and completes the restoration
process to form intact DNA circles. DNA ligase is responsible for joining
together fragments of DNA. In DNA replication, after the primers are replaced
by DNA Polymerase I, DNA ligase assists in the formation phosphodiester
bonds between the fragments. This is essential for creating one continuous
strand.

What role does Restriction Enzymes play during

replication?
Restriction enzymes (also known as restriction endonucleases) are proteins
which cut DNA up at specific sequences in the genome. They race along
strands of DNA and RNA looking for a specific sequence where it cuts and
renders that DNA harmless. The most common purpose is for security. If a
foreign molecule from a virus or somewhere else comes into a cell, it is cut
up to stop it from being transcribed. When these were discovered, scientists
used them for genetic engineering by opening a DNA molecule to insert a
gene.

Why is Cloning Important?

Molecular Cloning allows scientists to not only discover the what proteins are
present and their function, but also explore what happens in a cell when
these proteins are changed. When studying cell division, specifically,
scientists look for proteins that control the beginning and end of division.
Using the recombinant DNA (containing the both the human cell DNA and the
cloned plasmid), scientists can direct the replication within the human cells.
By manipulating cells with cloning and learning more about specific proteins,
scientists can take their research and apply it to larger-sale research
endeavors like diseases and pathogens.
It is important to note, however, that cloning is not used to study just
proteins involved in the cell cycle. Molecular cloning has led scientists to
discover the entire genetic sequences of many different species, inactivate
genes in both humans and other organisms, and create transgenic
organisms.

Construction of DNA Library

A DNA library is a collection of DNA fragments that have been cloned into
vectors so that researchers can identify and isolate the DNA fragments that
interest them for further study.
Below are the steps for creating a genomic library from a large genome.
1. Extract and purify DNA.
2. Digest the DNA with a restriction enzyme. This creates fragments that
are similar in size, each containing one or more genes.
3. Insert the fragments of DNA into vectors that were cut with the same
restriction enzyme. Use the enzyme DNA ligase to seal the DNA
fragments into the vector. This creates a large pool of recombinant
molecules.
4. These recombinant molecules are taken up by a host bacterium by
transformation, creating a DNA library.