geneticengineen_introduction_to enhance the production of protein inthe industial application_08_2024.pptx

geneticengineen_introduction_to enhance the production of protein inthe industial application_08_2024.pptx

1. BioE 6304 : Microbial Engineering

2. Genetic engineering: medicine, agriculture, industry Bacteriophages – structure, lifecycle, and isolation and culture of; manipulation of bacteriophages Expression of recombinant proteins in microbial systems - bacteria and yeast Pathway engineering and modification to produce secondary metabolites and enzymes in heterologous systems - (include bacteria, yeast, algae) Biofuel Microbial cell-surface engineering - modification of bacterial cell surfaces to interfere with host-microbial contact interactions; the functional anatomy of bacterial surfaces Our way in this course

3. S eminar presentations Assignments J ournal clubs C ase studies Test UE Assessment methods

4. Case Study 1 : Developing Microbial Strains for Bioremediation   Case Study 2: Microbial Production of Biofuels Case Study 3: Microbial Engineering for Pharmaceutical Production   Case Study 4: Engineering Probiotics for Human Health   Case Study 5: Microbial Engineering for Food Production C ase studies

5. GENETIC ENGINEERING

6. Any process by which genetic material is changed in such a way as to make possible of the production of new substances or new functions. Genetic Engineering

7. Prior 1950s Term “gene” was used to stand for the unit by which some genetic characteristics passed to generation. 1953 Englishc chemist Francis Crick & American biologist James Watson created the DNA structure Chemical Structure of Genes

8. Chemical Structure of Genes Are very long chains/units made up of a combination of simple sugar and phosphate group. DNA Attached to this chains are the nitrogen base ( A, T, C, G)

9. Chemical Structure of Genes Each DNA has a pattern CODONS (amino acids) Arranged into particular sequence protein

10. Is the process in which fragments of DNA from one or more different microorganism are combined to form rDNA (recombinant DNA) and are made to function within the cell of a host organism. 2 highly significant techniques: Gene transfer transferring the gene from one source to another subject. Gene therapy Correcting defective gene that are responsible for disease development. Gene Splicing

11. Gene Splicing

12. Plasmid - A circular form of DNA often used as a vector in genetic engineering. Vector – an organism/ chemical that is used to transport a gene to the host cell. Host cell – the cell into where the new gene is transplanted Enzymes used: Endonucleases – enzymes that cut DNA molecule at some given location Exonucleases – enzyme that removes one nitrogen base unit at a time Ligases – enzyme that join two DNA segments together Gene Splicing

13. Although the concept of gene transfer is relatively simple, its execution presents considerable technical obstacles.  American biochemist Paul Berg (1926-), often referred to as the “father of genetic engineering”. He developed a method for joining the DNA from two different organisms, a monkey virus known as  SV40  and a virus called  lambda phage. the American biochemists Stanley Cohen (1922-) at Stanford University, and Herbert Boyer (1936-) at the University of California and San Francisco, discovered an enzyme that greatly increased the efficiency of the Berg procedure.   Gene Transfer

14. INSULIN Produced by “ Genetech ”, first genetic engineering company, founded by Robert Swanson and Herbert Boyer. Obtains a copy of insulin gene (can be from natural source or manufactured) Inserting the insulin gene into the vector (using the gene splicing process) The hybrid plasmid can now be inserted to the host cell. ( this is the manufactured insulin that is injected to diabetic patients) Gene Transfer

15. Human growth hormone For children whose growth is insufficient bc of genetic problems Interleukin-2 For treatment of cancer Factor VIII Needed by hemophiliacs for blood clotting Erythropoietin For treatment of anemia Tumor necrosis factor For treatment of tumors Tissue plasminogen activator Use to dissolve blood clots Other rDNA products

16. 4 approaches A normal gene inserted to compensate for the defective gene. Abnormal gene replaced with a normal one Abnormal gene repaired through selective reverse mutation Change the regulation of gene pairs. Gene Therapy

17. Gene Therapy

18. A vector delivers the therapeutic gene into a patient’s target cell The target cells become infected with the viral vector The vector’s genetic material is inserted into the target cell Functional proteins are created from the therapeutic gene causing the cell to return to a normal state. Gene Therapy

19. The first gene therapy was performed on September 14 th 1990 Ashanti DeSilva was treated for SCID Doctors removed her white blood cells, inserted the missing gene into the WBC and then put them back into her blood stream It strengthened her immune system, but it only worked for a few months Gene Therapy

20. Genetic Engineering could increase genetic diversity, and produce more variant alleles which could also be crossed over and implanted into other species Another of genetic engineering is that diseases could be prevented by detecting people that are genetically prone to certain hereditary diseases, and preparing for the inevitable. As well as preventing disease, with genetic engineering infectious diseases can be treated by implanting genes that code for antiviral proteins specific to each antigen  Advantages

21. Animals and plants can be 'tailor made' to show desirable characteristics. Genes could also be manipulated in trees for example, to absorb more CO2 and reduce the threat of global warming.  Advantages

22. Nature is an extremely complex inter-related chain consisting of many species linked in the food chain. Some scientists believe that introducing genetically modified genes may have an irreversible effect with consequences yet unknown.  Genetic engineering borderlines on many moral issues, particularly involving religion, which questions whether man has the right to manipulate the laws and course of nature.  Disadvantages

23. Basic principle of Genetic Engineering DNA fragment of interest is obtained by cleaving chromosomes by Restriction endonuclease. Cloning vector is cleaved with Restriction endonuclease. Fragments are ligated to the prepared cloning vector. Recombinant vector DNA is introduced into the host cell Propagation (cloning) produces many copies of recombinant DNA The gene is extracted and harvested the product

24. Tools of Genetic Engineering Enzymes Restriction Endonucleases (REs) DNA ligase DNA Polymerases Reverse transcriptase Transformation factors Plasmid Bacteriophage Cosmid Yeast

25. Thanks for Listening