Name: Petra Franzén
Position: Laboratory Engineer
Gender: F
Actual starting date: 1 July, 1996
Administrative starting date: 1 July, 1996
Name: Eva Davey
Position: Laboratory Assistant
Gender: F
Actual starting date: 1 July, 1996
Administrative starting date: 1 July, 1996
In general this year has been spent planning the space that was available. The lab has been rebuilt and equipped, and we have also set up routines for how the work preferably should be performed. A list of the equipment available in the lab as well as short information and rules of how to use it will be distributed soon, together with our working routines.
We have also started to perform lab work ourselves as well as supervise PhD students and post-docs in molecular biology and protein expression work in various projects.
Petra Franzén has mainly been involved in planning of projects and practical guidance of people in various projects. Petra has for example been working with Mary-Rose Hoja in sub-cloning and recombinant expression of the extracellular immunoglobulin-like domains of the human PDGFa receptor, and with Jinyu Zou and Eva Davey in recombinant expression of the rat cytosolic epoxide hydrolase protein. A lot of time has also been spent to teach and inform visitors how to do DNA work as well as how to use the equipment in the lab e.g sonicator, PCR machine, electroporator, Bio-Rad minigel system etc. The first part of this year Eva has been working with Malin Björkman in sub-cloning and recombinant expression of a heme-binding lipoprotein, hbpA, from Haemophilus influenzae type b. Eva is currently working with in the epoxide hydrolase project, where she is going to sub-clone the rat cytosolic epoxide hydrolase protein into a vector that will give a fusion protein with glutathione S-transferase. The purpose with this is to increase the solubility of the desired protein and to simplify its purification. Moreover, Jan Saras has started as a post-doc and is currently working in the Expression lab. His project is focused on recombinant expression of the human membrane protein Band 4.1, which links the cytoskeleton to the plasma membrane.
In summary we can say that we are quite satisfied with the progress of the molecular biology and the protein expression techniques in the lab, but there are of course still a lot more that can, and will, be done.
Sometimes certain mammalian proteins are not soluble when they are expressed in bacterial systems. To overcome this problem one can either try to refold the insoluble protein or use another expressing system. Therefore, we have during the year started to investigate the possibility of establish a protein expression system in eukaryotic cells, i.e baculo expressing system. For this purpose we will need cell culture room for sterile work. We would also like to be able to do functional studies of the recombinantly expressed proteins in order to confirm that they are active and behave as the wildtype protein. This is what we will be focused on in the nearest future.
Latest update at 9 March, 1998.