Principles of Biotechnology and Genetic Engineering

Principles of Biotechnology and Genetic Engineering. Author: Dr. A.J. Nair; ISBN: ; Category: Engineering, Science, Biotechnology. ×.
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Enter your mobile number or email address below and we'll send you a link to download the free Kindle App. Then you can start reading Kindle books on your smartphone, tablet, or computer - no Kindle device required. Would you like to tell us about a lower price? If you are a seller for this product, would you like to suggest updates through seller support? Biotechnology is a fast developing technology as well as a science that has already made an impact to different aspects of day-to-day human life such as Public Health, Pharmaceuticals, Food and Agriculture, Bioenergetics and Information technology.

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It is very clear that biotechnology will be a key technology for the 21st century or the science of future. It has the potential to ensure food security, dramatically reduce hunger and malnutrition, and reduce rural poverty particularly in the developing countries like India.

Considering its commercial potential and its possible impact on the economy, Government of India has taken a number of measures to build up trained human resource in biotechnology and promote research and development and its commercial aspects in the country. This textbook covers all the fundamental aspects of Biotechnology. It has been written in a very simple manner explaining the fundamental concepts and techniques in detail so that it will be very easy to understand, even for those who do not have a basic understanding in biology.

Principles of Biotechnology and Genetic Engineering

Reading this textbook will give the learners an idea about the relationship between the subject and health, nutrition, agriculture, environment, industry etc. The book will be very helpful in developing interest in the beginners to study biotechnology as a discipline and pursue higher studies and research in biotechnology and other professional courses in future. Read more Read less. Save Extra with 1 offer. This specific base sequence is known as the Recognition Sequence for Hind Il. Restriction endonucleases recognizes a specific palindromic nucleotide sequence in DNA.

Palindromic nucleotide sequence is a sequence of base pairs thatreaads same on the two strands when orientation of the reading is kept the same. For example the following sequence reads the same on the two strands in 5' to 3' direction. This is also true when read in the 3' to 5' direction.

Principles of Biotechnology and Genetic Engineering : Dr. A. J. Nair :

This leaves the single stranded portions at the ends. These ends are called sticky ends because they form hydrogen bonds with complimentary cut counterparts. The DNA fragments separate according to their size through sieving effect of agarose gel. The separated DNA fragments are visualised by staining with ethidium bromide followed by exposure to UV radiation.


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The separated DNA fragments appear as bright orange coloured bands. The separated fragments of DNA are cut out from the agarose gel and extracted from the gel piece. This step is known as elution. Vectors are the vehicles which carry the foreign DNA into a given host cell. Salient features of a vector: The vector should contain an origin of replication Ori which facilitates the multiplication of vector within the host cell. The vector should have a selectable marker which allows to select and isolate the recombinants from non-recombinants.

The vector should contain atleast one unique restriction endonuclease recognition site to enable the insertion of foreign DNA into the vector.

The vector should be of smaller size which facilitates its entry into the host cells. The most efficient method of screening for the presence of recombinant plasmids is based on the principle that the cloned DNA fragment disrupts the coding sequence of a gene. This is termed as Insertional Inactiviation.

For example, the powerful method of screening for the presence of recombinant plasmids is referred to as Blue-White selection. This method is based upon the insertional inactivation of the lac Z gene present on the vector. The lac Z gene encodes the enzyme beta-galactosidase, which can cleave a chromogenic substrate into a blue coloured product. In order to attach the foreign DNA to a vector, the vector should have a recognition site for a specific restriction enzyme.

Multiple recognition sites will result in multiple DNA fragments, complicating the process of cloning. A vector has more than one antibiotic resistance gene. The foreign DNA is ligated into a restriction site in one of the antibiotic resistance genes. On ligating the foreign DNA into a recognition site within the tetracycline resistance gene, the plasmid loses its tetracycline resistance.

But, it can still be selected for non-recombinants by plating on ampicillin-containing medium. Now, by transferring the ones that grow in the ampicillin medium to a medium with tetracycline, we can dissect out recombinants from non-recombinants. The recombinants will grow in ampicillin but not in tetracycline medium; while non-recombinants will grow in both mediums. Currently, alternative markers are available that can differentiate recombinants from non-recombinants based on their ability to produce color.

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On reaction with a substrate, the recombinants do not produce color whereas non-recombinants produce color. Blue-white screening using B-galactosidase [Source: Long before us, bacteria and viruses knew how to transfer genes into plants and animals. These tumours then produce chemicals that the pathogen requires. With better understanding, we have now converted these pathogens into useful vectors to deliver genes of interest to plants or animals. This vector is no longer harmful but is useful in delivering genes of interest to plants.

Retroviruses transform normal cells into cancerous cells in animals. These have also been modified so that they are no longer harmful and can deliver genes to animals.


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Host cells are bacterial cells which take up the recombinant DNA.