Gene Cloning
To make it possible, we need the relevant protein in a large amount. It must be available continuously. So to treat humans animal proteins have been used. This can be done wherever possible but not all disorders can be treated with animal proteins. And some side effects have also been identified in patients like allergic symptoms to these proteins.
To produce animal proteins for human administration, a gene cloning process has been used. The best example is the insulin, Clotting Factor VIII, and interferon.
These proteins are produced by using gene cloning. Examples for these recombinant proteins include insulin, growth hormones, clotting factor VIII, interferon, etc.. To illustrate this process, let us now discuss briefly the process of the recombinant insulin production.
Transfer of insulin gene
The transfer is done with the help of a bacterial plasmid. It carries the gene into a replicating bacterial cell which in turn produces human insulin. Let us now discuss the process.
First by using the restriction enzyme the plasmid is cut across both strands. It makes loose, sticky ends. And the DNA can then be inserted.
To fit properly into the loose ends of the opened plasmid DNA ring, special linking sequences are added to the human cDNA. That is complementary DNA. The plasmid inserted with the human gene is termed as a recombinant plasmid can be now inserted to other organisms that are the bacterial cells being used. Now the bacterial cells and the plasmid vectors are mixed up in the organism.
Plasmids can make it entry into bacteria by the transfection process. When the bacteria have been inserted with the rDNA molecule it will lose its uniqueness of replication. They lose the replication capacity. Only the inserted Human cDNA will start multiplying and yield number of copies.
Cells divide very quickly you know. So the human cDNA will multiply into a millions of cell soon. We can have millions of clones containing the same human gene.
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