Insulin production by genetic engineering

presentation of insulin production by genetic engineering

1. Genetic engineering production of insulin and human hormones

2. HAWASSA UNIVERSITY 12/19/2018 2 Assignment presentation MSc : clinical laboratory ( special truck diagnosis and public health microbiology) Course : microbial genetics Lecturer : Dr. P. VIJAYALAKSHMI by : Abdikhaliq Hussein Ali

3. Learning Objectives At the end of this module, participants will be able to: Understand Definition, Function, Importance, History, of insulin Describe Insulin production using rDNA Technology Understand Definition, Function, Importance, History, of human growth hormone Describe HGH production using rDNA Technology Describe the genetic engineering of other human hormane production

4. Outline Introduction Structure and function of insulin Genetic engineering of insulin Structure and function of HGH Genetic engineering of HGH Genetic engineering of human hormones.

5. Introduction the earliest uses of biotechnology in pharmaceutical manufacturing is the use of  recombinant DNA  technology to modify Escherichia coli  bacteria to produce human  insulin , which was performed at  Genentech  in 1978 Insulin  is a hormone produced by β-cells of islets of Langerhans of pancreas. It was discovered by sir Edward Sharpey Schafer (1916) while studying Islets of Langerhans People who do not produce the necessary amount of insulin have diabetes.

6. Introduction

7. Structure of insulin Chemically, insulin is small and simple protein consist of 51 amino acids, 30 construct polypeptide chain B and 21 amino acids construct polypeptide chain A and both chains linked by disulfide bond

8. Genetic engineering for insulin production Microorganisms

9. Main process of insulin production Microorganisms

10. Process of Producing Insulin Using Recombinant DNA Technology

11. First Step (Preparing) The human gene is isolated. The mRNA is taken from the cell of islet of Langerhans. Messenger RNA is a molecule of RNA that encodes a chemical "blueprint" for a protein product. The isolated gene contains the code of the human DNA for the production of insulin. The plasmid DNA of the bacterial cell is taken out of the cell. NOTE : Escherichia coli (E. Coli) bacteria is widely used in producing insulin but yeast may also be used.

12. Second Step (Cutting) The plasmid DNA of the bacteria is cut out producing plasmid ring which is an empty segment of the DNA. A Restriction Enzyme is an enzyme that cuts DNA at specific recognition nucleotide sequences known as restriction sites. A segment of DNA known as sticky ends.

13. Third Step (Combining) With the plasmid ring open, the gene obtained from human cell that contains the code of protein responsible for the production of insulin is inserted into the plasmid ring and the ring is closed. The human insulin gene is now combined with the bacterial DNA plasmid. Mix the recombinant plasmid with bacteria.

14. Fourth Step (Inserting) Transformed : resulting DNA is inserted back to the bacteria .

15. Production The cells need nutrients in order to grow, divide , and live. While they live, the bacterial cell processes turn on the gene for human insulin and the insulin is produced in the cell. When the bacterial cells reproduce by dividing , the human insulin gene is also reproduced in the newly created cells.

16. Human growth hormone Growth hormone is one of the most important hormones in human body. Human growth hormone is secreted from somatotropic cells in the anterior pituitary gland . It is responsible for normal body growth and development of muscles, bones, lengthening of height etc. are all regulated by HGH

17. Structure of HGH The major isoform of the human growth hormone is a protein of 191 amino acids and a molecular weight of 22,124 daltons . The three-dimensional structure is stabilized by two disulfide bridges four helical structures.

18. Genetic engineering of HGH production

19. Oder steps would be used in laboratory engineer bacteria that could express the human gene coding for HGH Transform the rDNA in to an E. coli bacteria Isolate the mRNA of the HGH gene from human cell (pituitary gland cells) Generate cDNA the HGH gene using reverse transcriptase Insert the HGH cDNA gene into the bacterial vector E. Coli express the HGH

20. Lastly, scientists isolate the human growth hormone from the transformed bacteria. This human growth hormone is used to make the injection that is given to the patient. Simulate the isolation of the human growth hormone by removing the paper clips from the bag and placing them on the vials in the diagram on the right

21. Other human hormones of genetic engineering production

22. References T.A. BROWN ; GENE CLONING AND DNA ANALYSIS Sixth Edition published by Blackwell Publishing Ltd, 2010, © 2010, 2006 by T.A. Brown http:// www.onlinebiologynotes.com/human-insulin-production-by-genetic-engineering/ Glick, Bernard R. Molecular biotechnology : principles and applications of recombinant DNA / Bernard R. Glick, Jack J. Pasternak, and Cheryl L. Patten. — 4th ed . Hmge . Human insulin from second generation genetic engineering , Novo. Insulin, Grolier Electronic Publishing Inc., 1992; 234-237. https://nptel.ac.in/courses/102103013/35