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  • Biotechnology 09/11/09 G. Podgorksi, Biol. 1010
  • Biotechnology 09/11/09 G. Podgorksi, Biol. 1010
  • Biotechnology

    1. 1. BIOTECHNOLOGY “ Unwinding the helical mystery”
    2. 2. What is Biotechnology ??? <ul><li>Technology based on biological sciences that includes </li></ul><ul><li>Agriculture </li></ul><ul><li>Food science </li></ul><ul><li>Genetics </li></ul><ul><li>Medicine </li></ul>
    3. 3. Bioengineering <ul><li>Using scientific methods with organisms to produce new products or new forms of organisms </li></ul><ul><li>Techniques used to produce or modify plants and animals or its substances and utilize microorganisms for specific use </li></ul>
    4. 4. APPLICATIONS <ul><li>A multidisciplinary field involving number of interrelated subjects </li></ul><ul><li>SCIENCE </li></ul><ul><li>Physical sciences </li></ul><ul><li>Life sciences </li></ul><ul><li>Social sciences </li></ul><ul><li>MATHEMATICS </li></ul><ul><li>Statistics </li></ul><ul><li>APPLIED SCIENCES </li></ul><ul><li>Computer applications </li></ul><ul><li>Agriculture </li></ul><ul><li>Instrumentation </li></ul>
    5. 5. What Are the Stages of Biotechnology Development <ul><li>Ancient biotechnology - early history as related to food and shelter; Includes domestication </li></ul><ul><li>Classical biotechnology- built on ancient biotechnology; Fermentation promoted food production, and medicine </li></ul><ul><li>Modern biotechnology- manipulates genetic information in organism; Genetic engineering </li></ul>
    6. 6. Animal cloning Gene cloning for pharmaceutical production The promise and perhaps perils of embryonic stem cells DNA fingerprinting Genetically modified foods and the American-European opinion divide.
    7. 7. Basics of Biotechnology <ul><li>Biochemistry </li></ul><ul><li>Microbiology </li></ul><ul><li>Cell biology </li></ul><ul><li>Molecular biology </li></ul>
    8. 8. Areas of Biotechnology <ul><li>Genetic Engineering </li></ul><ul><li>Protein engineering </li></ul><ul><li>Bioinformatics </li></ul><ul><li>Immunology </li></ul><ul><li>Plant biotechnology </li></ul><ul><li>Animal biotechnology </li></ul><ul><li>Cancer biology </li></ul><ul><li>Environmental Biotechnology </li></ul><ul><li>Marine Biotechnology </li></ul><ul><li>Nano biotechnology </li></ul><ul><li>Pharmacology </li></ul>
    9. 9. GENETIC ENGINEERING <ul><li>Genetic modification of an organism’s genes </li></ul><ul><li>It uses techniques to alter the structure and characteristics of genes </li></ul>
    10. 10. Central dogma of Molecular Biology
    11. 11. Recombinant DNA technology <ul><li>Technology involving removing, modifying and inserting the gene of interest </li></ul><ul><li>rDNA technology requires </li></ul><ul><li>Vectors </li></ul><ul><li>Plasmids </li></ul><ul><li>Restriction Enzyme </li></ul><ul><li>DNA sequence and templates </li></ul><ul><li>Living organism </li></ul>
    12. 12. Producing Recombinant molecules <ul><li>Isolate the gene </li></ul><ul><li>Insert it in a host using a vector </li></ul><ul><li>Produce as many copies of the host as possible </li></ul><ul><li>Separate and purify the product of the gene </li></ul>
    13. 13. Plasmids are Used to Replicate a Recombinant DNA <ul><li>Plasmids are small circles of DNA found in bacteria </li></ul><ul><li>Plasmids replicate independently of the bacterial chromosome </li></ul><ul><li>Replication often produces 50-100 copies of a recombinant plasmid in each cell </li></ul><ul><li>Pieces of foreign DNA can be added within a plasmid to create a recombinant plasmid </li></ul>
    14. 14. Human Insulin
    15. 15. PCR <ul><li>Polymerase chain reaction is an in vitro technique used for the amplification of DNA </li></ul><ul><li>It can amplify 10^5 copies of DNA from two single strands of DNA after 30 cycles </li></ul><ul><li>This is achieved by Primers , complementary sequences of defined DNA templates </li></ul><ul><li>PCR technique is applied for detecting pathogens, study of DNA polymorphism, molecular mapping, DNA finger printing </li></ul>
    16. 16. PCR steps <ul><li>Denaturation at 94°c for 20-30 sec </li></ul><ul><li>Annealing at ~50°c for 20-60 sec </li></ul><ul><li>Extension at 72°c for 30-60 sec </li></ul>
    17. 17. PCR machine
    18. 18. Applications of Genetic Engineering <ul><li>Insertion of genes from one organism to another </li></ul><ul><li>Producing recombinant cytokines, growth factors, hormones like insulin, enzymes </li></ul><ul><li>Recombinant vaccines, nitrogen fertilizers, amylase synthesis for cellulose degradation using plasmid technology </li></ul><ul><li>Transgenic plants and animals </li></ul><ul><li>Animal cloning </li></ul><ul><li>DNA finger printing technology used in forensic sciences </li></ul>
    19. 19. Transgenic Animals Animals that undergo deliberate modification in the structure of its genome by inserting genes of other species. Transgenesis produced animals like mice, pig, cattle and sheep . This technology has applications in agriculture and medicine
    20. 20. Transgenic Plants <ul><li>Plants that can be genetically modified by inserting other plant genomes for the effective production of crops and plants using plant breeding or tissue culture techniques </li></ul><ul><li>This recombinant technology produced many insect, herbicide, fungal, bacterial and virus resistance varieties </li></ul>
    21. 21. Animal Cloning <ul><li>Animal Cloning is the process by which an entire organism is reproduced from a single cell taken from the parent organism and in a genetically identical manner. This means the cloned animal is an exact duplicate in every way of its parent; it has the same exact DNA </li></ul><ul><li>It can happen naturally for instance: Asexual reproduction in certain organisms and the development of twins from a single fertilized egg </li></ul>
    22. 22. Development of Cloning <ul><li>Scientists have been producing cloned animals from embryonic cells for long time but they do not live longer </li></ul><ul><li>The first cloned animal that lived longer and reproduced naturally later was, Dolly the sheep </li></ul><ul><li>Dolly was created from developed mammary gland cells taken from a full grown sheep </li></ul><ul><li>Pigs, cats, rats, horses and deer are successful cloned animals after dolly </li></ul>
    23. 23. Procedure of Animal cloning <ul><li>Extract DNA nucleus from developed embryonic cells </li></ul><ul><li>Implant into an unfertilized egg from which the nucleus had already taken </li></ul><ul><li>Cells are developed from the process of replication </li></ul><ul><li>These cells are implanted into host mothers </li></ul><ul><li>Cloned animal that resulted had a genetic make-up exactly identical to the genetic make-up of the original cell. </li></ul>
    24. 24. Reproductive cloning
    25. 25. The Next step of Cloning ?????
    26. 26. Extremely improbable
    27. 27. Bioinformatics <ul><li>Bioinformatics is the study of science in which biology , computer science and information technology merge into single field </li></ul><ul><li>Managing and analyzing biological data using advanced computing techniques </li></ul><ul><li>Major research efforts in the field include sequence alignment, finding gene, genome assembly, protein structure alignment, protein structure prediction, prediction of gene expression and protein-protein interactions, and the modelling of evolution </li></ul>
    28. 28. Computers and Bioinformatics <ul><li>Bioinformatics is the computer assisted data management field that helps us </li></ul><ul><li>Gather </li></ul><ul><li>Store </li></ul><ul><li>Analyze </li></ul><ul><li>Integrate </li></ul><ul><li>biological and genetic information ( data ) </li></ul><ul><li>Bioinformatics is the “Electronic Infrastructure of Molecular Biology ” </li></ul>
    29. 29. <ul><li>There are many different Bioinformatics tools available online that contains biological databases. Commercial software are also used by researchers </li></ul><ul><li>It is associated with massive databases of gene and protein sequences and structure/function information databases </li></ul><ul><li>New sequences, new structure, protein or gene function that are discovered, searched, gathered and deposited into databases </li></ul>
    30. 30. What is done ? <ul><li>Analysis and Interpretation </li></ul><ul><li>Development of new Algorithm and Statistics </li></ul><ul><li>Development and Implementation of tools </li></ul>
    31. 31. Units of Information <ul><li>DNA </li></ul><ul><li>RNA </li></ul><ul><li>Protein </li></ul><ul><li>Sequence </li></ul><ul><li>Structure </li></ul><ul><li>Evolution </li></ul><ul><li>Pathways </li></ul><ul><li>Interaction </li></ul><ul><li>Mutation </li></ul>
    32. 32. Biological Data used in Bioinformatics <ul><li>DNA ( Genome ) </li></ul><ul><li>RNA ( Transcriptome ) </li></ul><ul><li>Proteins ( proteome ) </li></ul>
    33. 33. DNA analysis <ul><li>Simple sequence analysis </li></ul><ul><li>Database searching </li></ul><ul><li>Pairwise analysis </li></ul><ul><li>Regulatory regions </li></ul><ul><li>Gene finding </li></ul><ul><li>Whole genome annotation </li></ul><ul><li>Comparative Genomics where species and strains can be analyzed </li></ul>
    34. 34. RNA analysis <ul><li>Splicing variant </li></ul><ul><li>Tissue specific expression </li></ul><ul><li>Structure </li></ul><ul><li>Single gene analysis ( various cloning techniques ) </li></ul><ul><li>Experimental data involving thousands of genes simultaneously </li></ul><ul><li>DNA chips, Microarrays and Expression arrays analysis </li></ul>
    35. 35. Protein <ul><li>Proteome of an organism </li></ul><ul><li>Structure </li></ul><ul><li>2D gel </li></ul><ul><li>2D structure </li></ul><ul><li>3D structure </li></ul><ul><li>Mass spectroscopy </li></ul>
    36. 36. <ul><li>01010111 01100101 01100101 01110100 00101101 </li></ul><ul><li>01 01 10 11 00 10 01 11 11 10 11 01 00 11 01 00 00 10 11 01 </li></ul><ul><li>A A C G T C A T T C G A T G A T T C G A </li></ul><ul><li>Just as we can teach a computer to predict things about a sequence of letters in English prose, we can also teach it to predict things about a other sequences —like a genetic sequence </li></ul>
    37. 37. Systems Biology <ul><li>Metabolic Pathways </li></ul><ul><li>Regulatory Networks </li></ul><ul><li>Whole organism Phylogeny </li></ul>
    38. 38. Applications <ul><li>Medical </li></ul><ul><li>Understand life processes in healthy and disease states </li></ul><ul><li>Genetic diseases </li></ul><ul><li>Biotech and Pharmaceutical </li></ul><ul><li>To find or develop new and better drug </li></ul><ul><li>Designing drug (Gene or Structure based) </li></ul><ul><li>Agriculture </li></ul><ul><li>Disease, drought, heat resistant plants </li></ul><ul><li>High yielding crops </li></ul>
    39. 39. DNA Microarray <ul><li>Gene expression patterns of whole genomic analysis of many organisms have been achieved by DNA microarrays . High density microarrays allowed global analysis of expression pattern to determine the cellular function of genes, the nature and regulation of biochemical pathways, and the regulatory mechanisms at play during certain signaling conditions or diseases. </li></ul>
    40. 40. Preparation of Microarrays
    41. 41. DNA Microarray
    42. 42. Applications of DNA Microarray <ul><li>Detect pathogens from blood samples </li></ul><ul><li>Examine targets for drug discovery </li></ul><ul><li>Determine gene expression changes for diseases like cancer </li></ul><ul><li>Identify inheritable markers which is used as genotyping tool </li></ul>
    43. 43. Cancer biology <ul><li>Cancer is a group of disease due to uncontrolled cell proliferation and differentiation </li></ul><ul><li>Cancer biology deals with studies of all types of cancer and different forms of therapies involved </li></ul>
    44. 44. How cancers are caused ? <ul><li>Any alteration in the genetic material </li></ul><ul><li>Effects of physical and chemical carcinogens </li></ul><ul><li>Errors in DNA replication or inheritance </li></ul><ul><li>Misexpressed growth factors </li></ul><ul><li>Activated Mutation and over expression </li></ul>
    45. 45. Cell Cycle
    46. 46. Apoptosis
    47. 47. Tumor suppressor genes <ul><li>Genes that reduces the probability of cells turning into tumor cells. </li></ul><ul><li>A mutation or deletion of such genes will increase the probability of formation of tumors </li></ul><ul><li>Examples : p53, RB1, WT1, DCC etc </li></ul><ul><li>Inactivation of one allele of some tumor suppressor genes is sufficient to cause tumors </li></ul><ul><li>Tumor markers are used to identify these genes. E.g. CA 27.29, PSA, CA-125 etc </li></ul>
    48. 48. Carcinogens ( Agents that cause cancer ) <ul><li>Physical carcinogenesis </li></ul><ul><li>Ionizing radiations like X-rays, α particles, γ rays which are >10eV </li></ul><ul><li>Non ionizing radiations like UV rays, microwaves, Ultrasound which are <10eV </li></ul><ul><li>Chemical carcinogenesis </li></ul><ul><li>Organic chemical carcinogen </li></ul><ul><li>Inorganic chemical carcinogen </li></ul><ul><li>Inert chemical carcinogen </li></ul><ul><li>Hormonal carcinogen </li></ul>
    49. 49. Types of cancer <ul><li>Generally cancers can be classified as Benign tumors and Malignant tumors </li></ul><ul><li>Carcinoma ( adenocarcinama, squamous cell carcinoma ) </li></ul><ul><li>Sarcoma ( fibro sarcoma, liposarcoma ) </li></ul><ul><li>Lymphoma ( chronic lymphocytic leukemia, AIDS related lymphoma ) </li></ul><ul><li>Blastoma (fibroma, chondroma, osteoma, adenoma ) </li></ul>
    50. 50. Screening and Diagnosis of cancer <ul><li>Screening test </li></ul><ul><li>Mammography </li></ul><ul><li>Cervical smear or Pap test </li></ul><ul><li>Endoscopies </li></ul><ul><li>Examination of faecal occult blood test </li></ul><ul><li>Diagnosis </li></ul><ul><li>Histopathological examination( biopsy ) </li></ul><ul><li>Endoscopy (broncho, colono, cysto, duodeno, gastro, lapro) </li></ul><ul><li>Biomarker studies (CA 15-3, PSA, AFP ) </li></ul><ul><li>Imaging ( x rays, MRI, CT, PET scan ) </li></ul>
    51. 51. Cancer therapy <ul><li>Chemotherapy </li></ul><ul><li>Radiation therapy </li></ul><ul><li>Gene therapy ( in vivo and ex vivo ) </li></ul><ul><li>Monoclonal antibody therapy </li></ul><ul><li>Immunotherapy </li></ul><ul><li>Hormonal therapy </li></ul><ul><li>Symptom control drugs </li></ul><ul><li>Angiogenesis inhibitor </li></ul>
    52. 52. Monoclonal antibodies <ul><li>Antibodies produced from single epitope of an antigen </li></ul><ul><li>Polyclonal antibodies that produces antiserum affect the normal cells </li></ul><ul><li>But MAbs produced from single clones react with single antigenic determinants </li></ul><ul><li>It’s a technique for in vitro production of antibodies using Hybridoma technology </li></ul>
    53. 53. Production of MAbs Fusing myeloma cells and spleen cells of mice
    54. 55. Uses of MAbs <ul><li>Antigen detection </li></ul><ul><li>Tumor related antigen detection </li></ul><ul><li>Imaging agent </li></ul><ul><li>Immunoscintigraphy </li></ul><ul><li>Pregnancy and Sex determination </li></ul><ul><li>Against viral and bacterial infections </li></ul><ul><li>Producing anti-idiotype antibodies </li></ul><ul><li>Immnuo diagnostic reagents </li></ul>
    55. 56. Plant Biotechnology <ul><li>Genetically modified crops and plants </li></ul><ul><li>Transgenic plants </li></ul><ul><li>Plant tissue culture </li></ul><ul><li>Therapeutic plants </li></ul><ul><li>Molecular pharming </li></ul><ul><li>Engineering plants genetically is Plant Biotechnology </li></ul>
    56. 57. Methods of plant gene delivery <ul><li>Agro bacterium mediated gene transfer (most widely used method) </li></ul><ul><li>Gene gun / biolistic method </li></ul><ul><li>Electroporation </li></ul><ul><li>Microinjection </li></ul><ul><li>Viral vector method </li></ul>
    57. 58. Ti plasmid of A.tumefaciens
    58. 59. Mode of gene transfer
    59. 60. Transgenic plants <ul><li>Genetically modified ( GM ) crops improved food production for the growing population by producing transgenic plants. GM crops include cotton, mustard, potato, maize, wheat, cassava, yams, sorghum, rice, millet etc. </li></ul>
    60. 61. GM technology <ul><li>Pest resistant plants </li></ul><ul><li>Herbicide resistant plants </li></ul><ul><li>Virus and fungal resistant plants </li></ul><ul><li>Modification of plant oil biosynthesis </li></ul><ul><li>Extended shell life of fruits </li></ul><ul><li>Medicinal plants producing vaccines </li></ul><ul><li>Nutritional enrichment/synthetic seeds </li></ul><ul><li>Tolerance to abiotic stress ( heat, drought, cold, saline ) </li></ul>
    61. 62. Transgenic rice <ul><li>Introduction of three genes into rice ( one from daffodil and two from microorganism ) exhibits an increased production of β -carotene as a precursor to vitamin A in the endosperm of the rice </li></ul>
    62. 63. Golden rice <ul><li>To alleviate vitamin A deficiency problem, development of a rice variety which produces β -carotene in the edible grains of the rice </li></ul>
    63. 64. Extended shelf life of fruit <ul><li>A major problem in fruit marketing is premature ripening and softening during transport. These changes are part of the natural ageing ( senescence ) process of the fruit. Ripening of fruit is closely associated with increase in polygalaturanase activity </li></ul>
    64. 65. Flavr Savr Tomato <ul><li>Antisense RNA against the enzyme activity developed to inhibit the synthesis of the enzyme and delayed the fruit ripening of tomato by GM technology. These tomato have longer shelf life called as “FLAVR SAVR TOMATO” </li></ul>
    65. 66. Plant tissue culture <ul><li>Plant tissue culture is a novel and innovative  technique to grow high quality, disease free plants quickly and in a large quantity by culturing and maintaining plant cells or organs like leaves, stem, root, branch shoot tip, petals, anther and pollen in sterile </li></ul>
    66. 67. Tissue culture process
    67. 68. Gene gun method
    68. 69. DNA finger printing
    69. 70. Basics of finger printing <ul><li>A DNA fingerprint is made by analyzing the sizes of DNA fragments produced from a number of different sites in the genome that vary in length </li></ul><ul><li>The more common the length variation at a particular site and the greater the number the sites analyzed, the more informative the fingerprint </li></ul>
    70. 71. Stem cell <ul><li>A stem cell is an undifferentiated, dividing cell that gives rise to a daughter cell like itself and a daughter cell that becomes a specialized cell type . </li></ul>
    71. 72. Embryonic stem cell
    72. 73. Nano biotechnology <ul><li>Nano-Technology is one of the emerging interdisciplinary fields which is about to bring a technological revolution. It is a engineering at the atomic or molecular scale, deals with devices typically less than 100 nanometers in size , or one billionth of a meter, or one ten-thousandth the width of a human hair </li></ul><ul><li>Nanotechnology provides a new basis for innovation in the life sciences, revolutionary biotechnology processes, the synthesis of new drugs and their targeted delivery, regenerative medicine, neuromorphic engineering stem cell research, genomics, proteomics as well as the well-established fields of agriculture, environmental management, medical device manufacturing </li></ul>
    73. 74. Nanobots <ul><li>A nanorobot is a specialized nanomachine designed to perform a specific task or tasks repeatedly and with precision. Nanorobots have dimensions on the order of nanometers </li></ul><ul><li>The nanorobot developed to kill the cancer cells would have a small computer, several binding sites to determine the concentration of specific molecules, and a supply of some toxin, which could be selectively released to kill a cell identified as cancerous. </li></ul>
    74. 76. Applications <ul><li>Nanobomb </li></ul><ul><li>Nanoimaging </li></ul><ul><li>Nano drug delivery </li></ul><ul><li>Microbivore </li></ul><ul><li>Artificial mechanical phagocytes </li></ul><ul><li>Tissue reconstruction </li></ul><ul><li>Frying tumors </li></ul><ul><li>Quantum dots </li></ul>
    75. 77. Ethical Arguments <ul><li>Ethical argument for or against the following topics </li></ul><ul><ul><li>Human Cloning </li></ul></ul><ul><ul><li>Gender Reversal on Livestock </li></ul></ul><ul><ul><li>Gender Reversal on Humans </li></ul></ul><ul><ul><li>Genetic Engineering of Plants </li></ul></ul><ul><ul><li>Genetic Engineering of Humans </li></ul></ul>
    76. 78. Career opportunities <ul><li>Biotechnology is a new and innovative industry with innumerable career options for anyone interested in life sciences. Biotechnology has changed the quality of life through improved medicine, diagnostics, agriculture and waste management . Because the field of biotechnology is still relatively new, there is an abundance of unique opportunities to make new discoveries and introduce useful innovations to the world. </li></ul>
    77. 79. Occupational opportunities <ul><li>Research </li></ul><ul><li>Quality control </li></ul><ul><li>Manufacturing and production </li></ul><ul><li>Regulatory affairs </li></ul><ul><li>Administration </li></ul><ul><li>Information system </li></ul><ul><li>Clinical research </li></ul><ul><li>Marketing and sales </li></ul>
    78. 80. Scope of biotechnology <ul><li>Genetically engineered microbes are today widely used for producing drugs and vaccines in large scale at low costs that are of great importance (human insulin, erythropoietin, and hepatitis-B vaccine) </li></ul><ul><li>Genetically engineering plants are also poised to produce vaccines </li></ul><ul><li>One of the future sources of cheap protein-drugs in the coming years, would be genetically engineered animals who would secrete drugs in abundance </li></ul>
    79. 81. The Indian advantage <ul><li>No other country in the world today has the unique set of advantages that India offers for large-scale practice of biotechnology. We have one of the largest bio in the world </li></ul><ul><li>We have at least seven distinct climatic zones and one of the largest and most varied sets of marine organisms anywhere </li></ul><ul><li>The ambient temperature in most parts of the country is just what living organisms need for their activities that result in a biotechnological product </li></ul><ul><li>We have large tracts of land available for growing the desired plants required for agriculture-based biotechnology </li></ul>
    80. 82. Future belongs to Biotechnology <ul><li>Biotechnology gives answer to many burning problems like food scarcity, ecological problems , pharmaceutical sector drop and spans its wings to almost all living World problems. Although there is lot of hype in media about the sector, but unfortunately in India [and in many countries surprisingly] it has not got due attention till yet. Government has started taking steps and new institutions are built and skill has started coming up </li></ul>