RECOMBINANT DNA TECHNOLOGY

Recombinant DNA technology :

Here is how recombinant technology works:

Recombinant technology begins with the isolation of a gene of interest. The gene is then inserted into a vector and cloned. A vector is a piece of DNA that is capable of independent growth; commonly used vectors are bacterial plasmids and viral phages. The gene of interest (foreign DNA) is integrated into the plasmid or phage, and this is referred to as recombinant DNA.


Before introducing the vector containing the foreign DNA into host cells to express the protein, it must be cloned. Cloning is necessary to produce numerous copies of the DNA since the initial supply is inadequate to insert into host cells.


Once the vector is isolated in large quantities, it can be introduced into the desired host cells such as mammalian, yeast, or special bacterial cells. The host cells will then synthesize the foreign protein from the recombinant DNA. When the cells are grown in vast quantities, the foreign or recombinant protein can be isolated and purified in large amounts.

Other uses for recombinant DNA

Recombinant DNA technology is not only an important tool in scientific research, but it has also impacted the diagnosis and treatment of diseases and genetic disorders in many areas of medicine. It has enabled many advances, including:

Isolation of large quantities of pure protein
In addition to the follicle-stimulating hormone (FSH) used in Follistim® AQ Cartridge and Follistim® AQ Vial, insulin, growth hormone and other proteins are now available as recombinant products. Physicians no longer have to rely on biological products (e.g. urine-derived FSH), that don't possess the same level of purity and consistency of recombinant products to treat their patients.


Identification of mutations
People may be tested for the presence of mutated proteins that may be associated with breast cancer, retino-blastoma, and neurofibromatosis.


Diagnosis of affected and carrier states for hereditary diseases
Tests exist to determine if people are carriers of the cystic fibrosis gene, the Huntington’s disease gene, the Tay-Sachs disease gene, or the Duchenne muscular dystrophy gene.


Transferring of genes from one organism to another
People suffering from cystic fibrosis, rheumatoid arthritis, vascular disease, and certain cancers may now benefit from the progress made in gene therapy.


Mapping of human genes on chromosomes
Scientists are able to link mutations and disease states to specific sites on chromosomes.

GOOD SITE FOR TOTAL MOLECULAR BIOLOGY:

1.http://www.molecular-plant-biotechnology.info/recombinant-DNA-technology/recombinant-DNA-technology.htm
2.http://www.biology.arizona.edu/molecular_bio/problem_sets/Recombinant_DNA_Technology/recombinant_dna.html



RECOMBINANT DNA TECHNOLOGY:

Recombinant DNA Technology - In nature, gene transfers are rather imprecise, and their range, in tenns of species involved, is remarkably limited. The above problems are circumvented by the recombinant DNA technology. A recombinant DNA molecule is produced by joining together two or more DNA segments usually originating from different organisms.

More specifically, a recombinant DNA molecule is a vector into which the desired DNA fragment has been inserted to enable its cloning in an appropriate host. This is achieved by using specific enzymes for cutting the DNA (restriction enzymes) into suitable fragments and then for joining together the appropriate fragments (ligation).


In this manner, a gene may be produced, which contains the. coding region from one organism joined to regulatory sequences from another organism; such a gene is called chimaeric gene. Clearly, the capability to produce recombinant DNA molecules has given man the power and opportunity to create novel gene functions to suit specific needs.

Recombinant DNA molecules are produced with one of the following three objectives:

(1) to obtain a large number of copies of specific DNA fragments,

(2) to recover large quantities of the protein produced by the concerned gene, or

(3) to integrate the gene in question into the chromosome of a target organism where it expresses itself. Even for the latter two objectives, it is essential to first obtain a large number of copies of the concerned genes.

To achieve this, the DNA segments are integrated into a self-replicating DNA molecule called vector; most commonly used vectors are either bacterial plasmids or DNA viruses. All these steps concerned with piecing together DNA segments of diverse origin and placing them into a suitable vector together constitute recombinant DNA technology.

The DNA segment to be cloned is called DNA insert

Recombinant DNAs are introduced into a suitable organism, usually a bacterium; this organism is called host, while the process is called transformation. The transformed host cells are selected and cloned.

The recombinant DNA present in such clones would replicate either in synchrony with or independent of the host cell; the gene present in 'the vector mayor may not express itself, i.e., direct the synthesis of concerned polypeptide. The step concerned with transformation of a suitable host with recombinant DNA, and cloning of the transformed cells is called DNA cloning or gene cloning.

However, often DNA or gene cloning is taken to include both the development of recombinant DNAs as well as their cloning in a suitable host. Similarly, often the term recombinant DNA technology is used as a synonym for DNA or gene cloning used in the broader sense. A rather popular term for these activities is genetic engineering.

A clone consists of asexual progeny of a single individual or cell, while the process/technique of producing a clone is called cloning. As a result, all the individuals of a clone have the same genotype, which is also identical with that of the individual from which the clone was derived.

Therefore, the genomes present in members of a single clone are also identical; this applies to the recombinant DNA as well. Therefore, gene or DNA cloning produces large numbers of copies of the gene/DNA being cloned.

Recombinant DNA Technology in the Synthesis of Human Insulin :

http://www.littletree.com.au/dna.htm

http://present.smith.udel.edu/biotech/rDNA.html.

SOME IMPORTANT QUESTIONS AND ANSWERS:

http://biog-101-104.bio.cornell.edu/BIOG101_104/tutorials/recomb_DNA.html

r DNA TECHNOLOGY BASIC TOOLS :

http://www.sci-ed-ga.org/modules/dna/index.html

Recombinant DNA Technology for the 21st Century:


http://www.biochemsoctrans.org/bst/034/0313/0340313.pdf

YOUTUBE BEST VIDEO ON rDNA TECHNOLOGY:

object width="425" height="355">