Recombinant DNA technology
Pharmaceutical Biotechnology Theory Notes
STEPS INVOLVED IN THE PROCESS
Isolate the gene of interest (foreign gene) from various sources for different applications. The cell or organism from which the required gene is taken is called a ‘donor’.
The gene is fragmented and isolated from the donor using restriction enzymes.
Adding the fragments with the desired gene into the ‘cloning vector’ (a plasmid, cosmid, or phage DNA), so as to develop recombinant DNA or chimeric DNA. Vectors are used to carry the desired gene from the donor to the hosts in which it will multiply.
The desired gene is inserted into the vector using suitable ligase enzymes which can be selected depending upon the type of bond to be formed between the foreign and vector DNA.
Introducing the recombinant vector into a suitable host cell in which it is able to multiply without interfering with the host functioning.
Replication or multiplication
Host has the ability to multiply and produce the required protein or any other substance for which it is formulated. Hosts are used for getting large quantity of product from the foreign gene which is our objective.
Overview Recombinant DNA technology
Application of genetic engineering in medicine
- Biotechnology plays an important role in the development of successful fermentation technology, producing recombinant proteins and peptides.
- These products have justified their uses as absolute supply, relative supply, safely specificity and economics.
- Naturally derived biological products, such as hGH, insulin, etc., of human origin are prone to contamination from their source.
- The rDNA products are safe as their use is concerned.
- In case of diagnostic products i.e. antibodies, the most significant aspect is specificity.
- The recombinant-derived proteins provide a higher level of sensitivity and specificity than had been available previously.
- Proteins are produced in minute quantity by living organisms, the genes for these have been identified, cloned and expressed at a high level in host cells.
- Production of insulin.
- Production of human growth hormone.
- Production of interferon.
SIMPLIFIED CONCEPTS OF BIOTECHNOLOGY By, Dr. Pramod Kadu & Ms. Suchita Vishwakarma
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