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Cloning
A major problem
in biochemical research is obtaining sufficient quantities of the
substance of interest. These difficulties have been largely
eliminated in recent years through the development of molecular
cloning techniques. The clone is a collection of identical
organisms that are all replicas of a single ancestor.
Methods of creating clones of desired properties, usually called
genetic engineering and recombinant DNA technology,
deserve much of the credit for the dramatic rise of biotechnology
since the mid-70'. The main idea of molecular cloning is to insert
a DNA segment of interest into an autonomously replicating DNA
molecule, called acloning vector, so that the DNA segment is
replicated with the vector. Such vectors could be, for instance,
plasmids (circular DNAs occuring in some bacteria). Reproduction
of DNA segments in appropriate hosts, results in the production of
large amount of the inserted DNA segment.
A DNA to be
cloned is usually a fragment of a genome of interest, obtained by
application of restriction enzymes. Most restriction enzymes
cleave duplex DNA at specific palindromic sites, generally two
fragments that have single strand ends that are complimentary with
each other (known as 'sticky ends'). Therefore, a restriction
fragment can be inserted into a cut made in a cloning vector by
the same restriction enzyme, because the segment ends stick
(chemically bond) to the loose ends of the vector. Such a
recombinant DNA molecule is inserted into a fast reproducing host
cell, and is duplicated in the process of the host's reproduction.
The cells containing the recombinant DNA are then isolated from
non-infected cells using an antibiotic substance which the
original vector is resistant to .
The cloning
technique provides both high quantities of DNA fragments, as well
as a mean to preserve them for long periods of time (by keeping
the host cells alive).
Next: The Human Genome Project
Up: Biological Technology
Previous: The Sequence Assembly problem
Itshack Pe`er
1998-12-27