These slides are from a workshop "Putting bioscience into context: exercises to enhance engagement" run at the Society for Experimental Biology conference in Canterbury (April 2006).

1. Dr Chris Willmott Dept of Biochemistry University of Leicester [email_address] Putting bioscience into context: exercises to enhance engagement Society for Experimental Biology (Canterbury) University of Leicester

6. Case study (2) – Wendy & Paul Wendy and Paul Carter have been married for twelve years. They would love to have children. Unfortunately, Wendy had breast cancer when she was 28 and although the chemotherapy has brought total remission from the disease it also caused damage to her ovaries that have made her infertile. Paul and Wendy have been on the waiting list at their local IVF clinic for a number of months awaiting donated eggs to try and have a baby. At present, however, there are 200 potential mothers seeking each donated egg and the couple know that realistically they may never receive a donated egg via the normal channels. Researchers at the hospital attached to the IVF clinic have recently gained permission to carry out experimental procedures using eggs harvested from aborted foetuses. The technique is controversial, but for Paul and Wendy it may represent their only chance to receive a donated egg. University of Leicester

7. What are the issues involved in this case? - Feel free to include aspects of the case that are likely to be issues for other people, your contributions need not be limited to your own opinions. Case study (2) – Wendy & Paul University of Leicester

10. Pharmaceutical development (1) Concerned by news coverage of increasing MRSA infection rates, you and your fellow directors of SmartaPharma plc decide to develop a new antibacterial compound. What stages will you need to go through from your initial idea until you have an effective (and profitable) medicine available for patients? University of Leicester

17. © Annette Cashmore, GENIE CETL, University of Leicester Card-based problems (1) Production of Medically Important Proteins Using Recombinant DNA Techniques

18. 8 © Annette Cashmore, GENIE CETL, University of Leicester Card-based problems (2) FLOW DIAGRAM OF CLONING A GENE You must now decide how to clone your gene : (1) cDNA library - Go to 9 (2) genomic library - Go to 10

19. 36 © Annette Cashmore, GENIE CETL, University of Leicester Card-based problems (3) YEAST VECTORS YEp213 Multicopy, DNA plasmid Shuttle vector - bacterial and yeast origins of replication (ORI) 2µm sequences (ORI andSTB) Selective markers : Yeast : auxotrophic marker - LEU2 gene (LEU2) Bacteria : Ampicillin resistance (Amp R) Tetracycline resistance (Tet R) Cloning vector Go to 37

20. YEAST-SUMMARY You have cloned your gene into a yeast expression vector. Check the table to see if you have expressed a protein. 76 © Annette Cashmore, GENIE CETL, University of Leicester Card-based problems (4) 78 yes Somatostatin genomic Hind III pJP31 78 yes Factor IX 78 yes hEGF 78 yes Somatostatin cDNA Hind III pJP31 81 no Any cDNA / genomic Bam HI YEp213 Go to Expression Protein Insert DNA Enzyme site Plasmid

21. 81 FAILURE !!!!! You have not been able to express your gene in this host - vector system. The reasons for this are : You have cloned into pBR322 or YEp213. These are cloning vectors which have no expression signals, also the insert DNA does not have any expression signals therefore there is no expression of the protein. Choose an expression vector - Go to 34, 36 or 38. N.B . Another possible reason for the absence of expression is that the gene may have been inserted into the vector in the incorrect orientation, therefore is unable to use the vectors promoter sequences. If the sequence or restriction map of the gene is known then restriction mapping can be used to determine whether this is the case. © Annette Cashmore, GENIE CETL, University of Leicester Card-based problems (5)

22. 83 SUCCESS Well done !!!! You have successfully produced your protein and it is biologically active. Producing a cell that synthesises large amounts of a desired protein is only the first stage in achieving a useful process. It is important to be able to recover the protein by a simple, economical method that results in high yields of a biologically active protein. Cell cultures can be scaled up and grown in large fermentors, and then the protein can be purified using a number of methods. © Annette Cashmore, GENIE CETL, University of Leicester Card-based problems (6)

29. Cancer biology Cell cycle control H uman e pidermal growth factor r eceptor (HER2) Monoclonal antibodies Therapeutic antibodies Resource allocation Clinical trials Herceptin

30. Therapeutic cloning Research ethics Science of stem cells Fraud Ethics of stem cells research Hwang cloning scandal

31. Mode of action (Neuraminidase inhibitors) Should wild birds be culled? Medicines: Tamiflu (Oseltamivir), Relenza (Zanamivir) Public perception of risk Bird flu Virus biology Limited drugs/vaccines: who gets them first?