Desarrollo de nuevos marcadores de selección para células de mamífero


Jaroslavsky Dodo, Ariel


In the production of biological molecules, it is convenient to have biocatalysts that express the product of interest in a stable way. For this, selection markers are used, which allow the selection of cells that have been modified and incorporated the genetic material that codes for a protein of interest, as well as maintaining the modified cell population. BHRF1 is a Bcl-2 homologous protein from the Epstein-Barr virus. After viral infection, cells activate the apoptotic cascade.By interacting with pro-apoptotic proteins, BHRF1 can repress the intrinsic pathway of apoptosis, lengthening the stay of the host in the cell. This assignment proposes to use this protein as a selection marker for mammals, using conditions that induce the intrinsic pathway of programmed cell death. This has a certain degree of economic relevance, given the posibility to save on antibiotics to select mammalian cultures. There is also scientific relevance it as it provides a new alternative to select transfected cells.
An experimental design was developed to find out the conditions that allow BHRF1 to act as a selection marker, as well as to evaluate the efficiency of this selection and the effect of the presence of this protein on the production of others. This approach brings up the need to build 3 plasmids: pGFPneo3, GFP-BHRF1 and BHRF1puro3. pGFPneo was build and sequencied. A reasonable candidate for GFP-BHRF1 was obtaind but not sequenced.



Lecina Veciana, Martí
Leivar Rico, Pablo 


IQS SE - Undergraduate Program in Biotechnology