LECTURE 1-1.pdf

INTRODUCTION TO BIOTECHNOLOGY BY DR AISHA MOHAMMED COURSE CONTENT INTRODUCTION TO BIOTECHNOLOGY TECHNIQUES IN BIOTECHNOLOGY SUCH AS CUTTING AND JOINING OF DNA MOLECULE,CLONING,POLYMERASE CHAIN REACTION(PCR) BASIC TOOLS USED IN BIOTECHNOLOGY INTRODUCTION TO BIOINFORMATICS...

INTRODUCTION TO BIOTECHNOLOGY BY DR AISHA MOHAMMED COURSE CONTENT INTRODUCTION TO BIOTECHNOLOGY TECHNIQUES IN BIOTECHNOLOGY SUCH AS CUTTING AND JOINING OF DNA MOLECULE,CLONING,POLYMERASE CHAIN REACTION(PCR) BASIC TOOLS USED IN BIOTECHNOLOGY INTRODUCTION TO BIOINFORMATICS INTRODUCTION TO BIOTECHNOLOGY  These products range from agricultural products (genetically modified foods) , pharmaceutical and medicinal products (vaccines and monoclonal antibodies) , industrial raw materials (synthetic raw materials), military products (bio-hazard weapons) There are various definitions of biotechnology depending on the applications of the technology AREAS OF BIOTECHNOLOGY (Colour codes)  White Biotechnology – Industrial Biotechnology, primarily bio catalysis and the production of chemicals/materials from renewable resources, as well as (hazardous) waste treatment  Red Biotechnology – Medical and pharmaceutical biotechnology  Green Biotechnology – Agricultural biotechnology, primarily genetic engineering of plants  Blue Biotechnology – Marine and aquatic applications of biotechnology, e.g. fish farming and algae cultivation  Grey biotechnology- conservation and restoration of contaminated natural ecosystems  Gold biotechnology- also known as bioinformatics; responsible for obtaining, storing, analyzing and separating biological information especially that related to DNA and amino acid sequences TIMELINE OF BIOTECHNOLOGY ANCIENT BIOTECHNOLOGY-early history as related to food and shelter, includes domestication CLASSICAL BIOTECHNOLOGY-built on ancient biotechnology; fermentation promoted food production and medicine MODERN BIOTECHNOLOGY-manipulates genetic information in organisms, Genetic engineering TYPES AND APPLICATIONS OF BIOTECHNOLOGY Microbial Biotechnology Agricultural Biotechnology Animal Biotechnology Forensic Biotechnology Bioremediation Aquatic Biotechnology Medical Biotechnology Pharmaceutical Biotechnology Pharmaceutical Biotechnology Biopharmaceutical drugs generated through researches in cell biology, genetics and recombinant DNA technology. Recombinant DNA (rDNA) and monoclonal antibody (mAb) are providing exciting opportunities for new pharmaceuticals development as well as new approaches to drug delivery During the last decade, over 359 biotechnology products were developed by more than 40 pharmaceutical companies for indications ranging from haemophilia, sepsis, skin ulcers, and rheumatoid arthritis to cancer. More than one-third of all biotechnology products are geared toward cancer therapy Biotechnology derived drugs are being applied in cancer therapy, HIV, AIDS and AIDS-related and autoimmune diseases In diagnostic investigations such as blood substitutes, clotting factors, etc. Products of Biotechnology  Antibiotics  Vaccines  GM foods  Weapons  Surfactants  Insecticides  Clothing Material  Diagnostic tools  Industrial Raw materials  Bioplastics FIELDS OF BIOTECHNOLOGY Genetics Cell Biology Bioinformatics Genomics Molecular Biology Proteomics Biophysics Metabolomics Synthetic Biology PROS OF BIOTECHNOLOGY It can improve health and reduce hunger simultaneously  Our food supply now contains more nutrients because of biotechnology.  Croplands can create essential vitamins and minerals, which decreases health problems caused by nutrient deficiencies.  People can consume less and still get the same nutritional benefits thanks to biotechnology  Which also increases farmland yields and nutritional richness. Consequently, more individuals can obtain the food they require.  It creates flexibility within the food chain.  Croplands can produce food with the use of biotechnology that might not be achievable under "normal" circumstances.  It is feasible to cultivate crops in the desert by using theories from this field of study.  Crops that are naturally pest-resistant can be developed.  Despite the fact that the amount of land our planet can support is limited, biotechnology enables us to use more of it for our needs.  It offers medical advancement opportunities.  Biotechnology allows us to look within just as easily as we can look to the outside world for advancement.  Studies that involve the human genome have allowed us to understand more about genetic diseases and some cancers, creating more effective treatments for them – and sometimes cures.  It has allowed us to explore the reasons behind certain birth defects to understand the importance of folic acid. That makes it possible to extend average human lifespans  It allows us to preserve resources.

 Biotechnology gives us an opportunity to extend the lifespan of our food supplies.  Practices that include salting foods to preserve them which date back beyond Biblical times.  Freezing and drying foods as methods of preservation have been known for centuries.  Pasteur pioneered an approach of heating food products to remove harmful elements so they can be preserved for an extended period.  It helps us minimize or eliminate waste products.  Biotechnology allows us to create waste products that have better biodegradable properties.  It allows us to manage landfills more effectively.  That way we can begin to minimize the footprint being left for future generations. It can reduce infectious disease rates.

 Biotechnology has helped us to create vaccines.  It has helped us be able to create treatments that reduce difficult symptoms of disease.  It has even helped us to learn how infectious diseases can be transmitted so their transmission can be reduced.  That allows us to protect those who are most vulnerable to these diseases, giving them a chance to live a happy, fulfilling life CONS OF BIOTECHNOLOGY It creates an all-or-nothing approach.

 One of the biggest problems that biotechnology faces is a lack of genetic diversity.  The processes included in this field can increase crop yields and improve medical science, but it comes at the price of a genetic bottleneck.  Should something unforeseen happen, an entire crop or medical treatment opportunity could go to waste or even threaten the survival of certain species.  It is a field of research with many unknowns.

 Although our database of biotechnology has greatly expanded in the last generation, there are still many long-term unknowns that we face.  What happens if we tinker with the genetics of a person to treat a disorder?  What happens to the environment if we dramatically alter crops to grow in locations that would normally not support crop growth?  Should every action have an equal and opposite reaction, future generations could pay the price for our research that is happening today.  It could ruin croplands.

 Biotechnology has allowed more vitamins and minerals to enter our food chain, but it could be coming at a cost.  Many crops obtain their nutritional content from the soil in which they grow.  If that soil is overloaded by the crop, it may lose its viability, even with crop rotation occurring.  That may reduce the amount of growing time each land segment is able to provide while extending its recovery period at the same time. In some situations, the croplands could be permanently ruined  It can be used for destruction.

 All the benefits that biotechnology can provide could also be turned into a weapon that is used for mass destruction.  Crops can be improved, but they can also be destroyed.  Medicines can be made with biotechnology, but diseases can also be weaponized.  If left unchecked, biotechnology could even create a societal class that is created specifically for research purposes only BASIC TOOLS AND TECHNIQUES USED IN BIOTECHNOLOGY RECOMBINANT DNA TECHNOLOGY RDT (GENETIC ENGINEERING)  Recombinant DNA technology involves the group of techniques used to cut up and join together genetic material, especially DNA from different biological species, and to introduce the resulting hybrid DNA into an organism in order to form new combinations of heritable genetic material.  Three types of biological tools are used in synthesis of recombinant DNA. These are as follows: 1. Enzymes:  cleaving enzymes (e.g., exo-nucleases, endonucleases, restriction endonucleases; exonuclease removes nucleotides from the end of the DNA while endonuclease cuts at specific positions within the DNA)  synthesizing enzymes (e.g., reverse transcriptase)  joining enzymes (e.g., ligases)  alkaline phosphatases 2. Vector or Vehicle DNA: The DNA used as a carrier for transferring a fragment of foreign DNA into a suitable host is called vehicle DNA or cloning vector or gene carrier (e.g., plasmids, bacteriophage DNA, cosmid – an artificial plasmid, phagemid/phasmid, artificial chromosome vectors). 3. Passenger DNA: It is the DNA which is transferred from one organism into another by combining it with the vehicle DNA (e.g., complementary DNA- cDNA, Synthetic DNA— sDNA, random DNA) Enzymes used in recombinant DNA technology
1. DNA ligase 2. Reverse transcriptase 3. Restriction endonuclease 4. Terminal transferase 5. DNA polymerase 1. DNA ligase: DNA ligase is isolated from E.coli infected with a lytic Bacteriophage T4 and used in recombinant DNA technology. The enzyme DNA ligase joins the DNA fragments with cloning vector. 2. Reverse transcriptase: RT is used to synthesize complementary strand (cDNA) from mRNA template. It is also known as RNA dependent DNA polymerase It is isolated from retrovirus 3. Restriction endonuclease: Restriction endonuclease enzyme recognize and cut DNA strand at specific sequence called restriction site. These enzyme is isolated from wide variety of microorganisms. There are 3 types of restriction endonuclease:  Type I Restriction endonuclease: It has both methylation and endonuclease activity. It require ATP to cut the DNA It cuts DNA about 1000bp away from its restriction site eg. EcoKI Type II Restriction endonuclease: It does not require ATP to cut DNA It cuts DNA at restriction site itself eg. EcoRI, Hind III Type III Restriction endonuclease: It requires ATP to cut DNA It cuts DNA about 25bp away from restriction site. eg. EcoPI 4. Terminal transferase: It is the enzyme that converts blunt end of DNA fragments into sticky end. If the restriction enzyme cuts DNA forming blunt ends, then efficiency of ligation is very low. So the enzyme terminal transferase converts bunt end into sticky end. Terminal transferase enzyme synthesize short sequence of complementary nucleotide at free ends of DNA, so that blunt end is converted into sticky end. 5. DNA polymerase: DNA polymerase is a complex enzyme which synthesize nucleotide complementary to template strand. It adds nucleotide to free 3′ OH end and help in elongation of strand It also helps to fill gap in double stranded DNA. DNA polymerase-I isolated from E. coli is commonly used in gene cloning Taq polymerase isolated from Thermus aquaticus is used in PCR HAZARDS OF GENETIC ENGINEERING Spread of New Diseases: New dangerous forms of microorganisms can be developed through recombinant DNA technique either accidentally or deliberately. Escape of such microorganisms from the research laboratory through drainage, laboratory glassware, laboratory personnel etc., may lead to the spread and origin of new diseases, which may pose a serious problem. HIV (AIDS Virus) is believed to be one such product Effect on Evolution: Nature has provided several barriers for exchange of DNA between prokaryotes and eukaryotes. Recombinant DNA technology permits exchange of DNA between these two classes of organisms and thus interferes with the natural process of evolution  Biological Warfare: There is always a possibility that some unscrupulous countries may use genetic engineering technology for biological warfare. In such warfare, disease-carrying microorganisms can be used against the enemy. This is bound to lead to disaster. 
Therapeutic cloning
 This the development of human embryos for use in research.  These embryos do not develop to fully grown individuals, rather, they are destroyed after five days when they are at the blastocyst stage.  At this stage stem cells are harvested and used to develop various organs for medical research.  Therapeutic cloning enables the cultivation of stem cells that are genetically identical to a patient  The stem cells could be stimulated to differentiate into any of the more than 200 cell types in the human body  The differentiated cells then could be transplanted into the patient to replace diseased or damaged cells without the risk of rejection by the immune system  These cells could be used to treat a variety of conditions, including Alzheimer disease, Parkinson disease, diabetes mellitus, stroke, and spinal cord injury  Both reproductive cloning and therapeutic cloning make use of Somatic Cell Nuclear Transfer