Thursday, January 23, 2014

The Process Of Molecular Cloning


by Armand Zeiders


Molecular cloning refers to a process where the DNA within a cell is reproduced or replicated. Scientists use molecular cloning in several ways, mainly to learn more about proteins within cells and the function of these proteins. In addition, scientists also study what happens when proteins are changed. The results of these studies are often used to fight diseases and for other medical discoveries.

The first step in the process of molecular cloning is to isolate the DNA from the cell that contains the actual gene you wish to replicate. You also must choose a host organism and cloning vector, which is a tiny piece of DNA into which a fragment of different DNA can be placed. Typically vectors are taken from viruses or plasmids, and these vectors are called recombinant DNA.

After you have isolated your DNA, it will need to be purified and then fragmented. This is done using what is known as a restriction enzyme. What these restriction enzymes do is generate staggered cuts in precise sequences, and these in turn will produce fragments with endings that are cohesive.

Once these fragments have been created, they are placed into the cloning vectors, which are the structures inside of our cells that can reproduce independently. In molecular cloning, the plasmids that are used have only one restriction site. When this site is split into fragments with a restriction enzyme, then those all-important cohesive endings have been created. Amazingly, these ends are identical to the initial DNA fragments.

Now that these fragments line up exactly with your plasmid, they must be bonded together. An enzyme known as DNA ligase is used as a glue or bond to hold together these fragments. Once the bonding has occurred, they can be placed into cells where they will begin to replicate, and identical copies of the DNA you have created will be produced. Then the host cell will divide, and your gene will be replicated again. As the cells begin to divide further, a cluster of cells with the new gene is created and now can be studied or used in various ways.

Most of the time, researchers actually contract out the work of molecular or gene cloning. There are companies that can provide everything from recombinant DNA production to gene cloning and even mutagenesis, which is the mutating of genes. Genes are often mutated so that researchers can study the altered DNA sequences and better understand the function of an amino acid or nucleotide in a certain protein.




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