What do you learn from this course?

Introduction to RDNA.

Definition

Steps

Applications

Limitations

Introduction:

Recombinant DNA technology is the joining together of DNA molecules from two different species.

The recombined DNA molecule is inserted into a host organism to produce new genetic combinations.

Why we need this and what is the purpose in daily life scenarios?

Its add value to science, medicine, agriculture and industry solutions.

The idea of recombinant DNA was first proposed by Peter Lobban, a graduate student of Prof. Dale Kaiser in the Biochemistry Department at Stanford University Medical School.

Stanford University applied for a US patent on recombinant DNA in 1974, listing the inventors as Herbert W. Boyer (professor at the University of California, San Francisco) and Stanley N. Cohen (professor at Stanford University); this patent was awarded in 1980.[

Definition:

“the joining together of DNA molecules from different organisms and inserting it into a host organism to produce new genetic combinations that are of value to science, medicine, agriculture and industry.”

A series of procedures that are used to join together (recombine) DNA segments. A recombinant DNA molecule is constructed from segments of two or more different DNA molecules.

It is the technology to produce an artificial DNA molecule by combining two or more fragments of DNA which are not necessarily associated with each other.

STEPS:

Steps for recombinant DNA technology. Bacterial plasmids with a resistance gene present (usually ampicillin or kanamycin resistant) are used as vector DNA.

(1) Isolate the DNA sequence gene of interest.

(2) Insert your DNA of interest into the vector plasmid: (a) cut both vector DNA and gene of interest with the same restriction enzymes, (b) mix the DNAs: they join by base pairing, (c) add DNA ligase to bond the pieces covalently. The result is a recombinant plasmid.

(3) Put recombinant plasmid into E. coli by transformation.

(4) Clone cells: (a) plate cells onto agar containing ampicillin (or kanamycin), (b) each colony contains the recombinant plasmid by their ability to grow in the presence of ampicillin in the agar.

(5) Pick colonies and inoculate a large culture to expand the plasmid carrying the gene of interest.

(6) Isolate and clean the DNA from the expanded culture.

(7) Transfer the plasmid DNA to desired cells (e.g., bacteria, yeast, mammalian cells, etc.) and test the biological properties of the gene of interest.

1. Production of Transgenic Plants:

By utilizing the tools and techniques of genetic engineering it is possible to produce transgenic plants or the genetically modified plants. Many transgenic plants have been developed with better qualities like resistance to herbicides, insects or viruses or with expression of male sterility etc.

2. Production of Transgenic Animals:

By the use of rec DNA technology, desired genes can be inserted

into the animal so as to produce the transgenic animal. The method of rec DNA technology aids the

animal breeders to increase the speed and range of selective breeding in case of animals. It helps for the

production of better farm animals so as to ensure more commercial benefits.

Another commercially important use of transgenic animals is the production of certain proteins and

pharmaceutical compounds. Transgenic animals also contribute for studying the gene functions in

different animal species. Biotechnologists have successfully produced transgenic pigs, sheep, rats and

cattle.

•5. Biosynthesis of Interferon:

Interferon’s are the glycoproteins which are produced in very minute

amounts by the virus-infected cells. Interferon’s have antiviral and even anti-cancerous properties. By

recDNA technology method, the gene of human fibroblasts (which produce interferon’s in human

beings) is inserted into the bacterial plasmid.These genetically engineered bacteria are cloned and

cultured so that the gene is expressed and the interferon’s are produced in fairly high quantities. This

interferon, so produced, is then extracted and purified.

6. Production of Antibiotics:

Antibiotics produced by microorganisms are very effective against different

viral, bacterial or protozoan diseases. Some important antibiotics are tetracyclin, penicillin,

streptomycin, novobiocin, bacitracin, etc.recDNA technology helps in increasing the production of

antibiotics by improving the microbial strains through modification of genetic characteristics.

7. Production of Commercially Important Chemicals:

Various commercially important chemicals can be

produced more efficiently by utilizing the methods of rec DNA technology. A few of them are the

alcohols and alcoholic beverages obtained through fermentation; organic acids like citric acid, acetic

acid, etc. and vitamins produced by microorganisms.

•8. Application in Enzyme Engineering:

•As we know that the enzymes are encoded by genes, so if there are

•changes in a gene then definitely the enzyme structure also changes. Enzyme engineering utilizes the

•same fact and can be explained as the modification of an enzyme structure by inducing alterations in the

•genes which encode for that particular enzyme.

•9. Prevention and Diagnosis of Diseases:

•Genetic engineering methods and techniques have greatly solved

•the problem of conventional methods for diagnosis of diseases. It also provides methods for the.

•prevention of a number of diseases like AIDS, cholera, etc. Monoclonal antibodies are useful tools for

•disease diagnosis. Monoclonal antibodies are produced by using the technique called hybridoma

•technology.

•10. Gene Therapy:

•Gene therapy is undoubtedly the most beneficial area of genetic engineering for human

•beings. It involves delivery of specific genes into human body to correct the diseases. Thus, it is the

•treatment of diseases by transfer and expression of a gene into the patients’ cells so as to ensure the

•restoration of a normal cellular activity.

•11. Practical Applications of Genetic Engineering:

•recDNA technology has an immense scope in Research

•and Experimental studies.

•It is applied for:

•a. Localizing specific genes.

•b. Sequencing of DNA or genes.

•c. Study of mechanism of gene regulation.

•d. Molecular analysis of various diseases.

•e. Study of” mutations in DNA, etc.

•12. Applications in forensic science:

•The applications of rec DNA technology (or genetic engineering) in

•forensic sciences largely depend on the technique called DNA profiling or DNA fingerprinting. It

•enables us to identify any person by analyzing his hair roots Wood stains, serum, etc. DNA

•fingerprinting also helps to solve the problems of parentage and to identify the criminals.

•13. Biofuel Production: Biofuels are derived from biomass and these are renewable and cost effective.

•Genetic engineering plays an essentially important role in a beneficial and largescale production of

•biofuels like biogas. bio hydrogen biodiesel bio-ethanol., etc. Genetic engineering helps to improve

•organisms for obtaining higher product yields and product tolerance.

•14. Genetically stable high producing microorganisms are being developed by using modern recDNA

techniques, which aid in an efficient production of bioenergy.

15. The energy crop plants are those plants which use solar energy in a better way for production of

biomass. Genetic improvements of these energy crop plants greatly help for quick and high Product on

of biomass which in turn reduces the biofuel production cost. The fermenting microbes which are

utilized for biogas production are improved at the genetic level for achieving better result.

16. Environment Protection:Genetic engineering makes its contributions to the environment protection in

various ways. Most important to mention are the new approaches utilized for waste treatments and

bioremediation Environment protection means the conservation of resources and hence to limit the

degradation of environment.