Volume IV, 2004
Dr. Michael Lentz
The immediate objective of our research aims to create stable cell lines to generate sufficient amounts of viral E1. The pMSG vector was chosen for its ability to amplify gene expression by way of hormonal or chemical induction, and for its ability to be selected under ampicillin medium by way of the Amp gene. pMSG was ligated with E1 insert in two fashions, blunt or modified ends. Inserts were created by way of digesting them with BamH1. The inserts were blunted and ligated with a blunt ended pMSG, or blunted, modified, digested with Sal 1 and then ligated with Sal 1 digested pMSG. Recombinants were transformed in E. coli cells, plated in ampicillin medium, and analyzed by way of electrophoresis. When analyzing the blunt ended technique, bacterial contamination was depicted by faint bands in the upper region of the gel. Products could not be analyzed from the Sal 1 ligated technique, due to complications and errors in plating. reating sufficient amounts of E1 protein by way of using blunt and modified ends to form recombinants proved difficult, yielding no desired products. Therefore, there must be further research to find a protocol to generate sufficient amounts of E1 from mammalian cell lines.
Orallo, Peaches Marie Raval, "Analysis of Papilloma Virus E1 Protein: Creation of Stable Cell Lines to Generate Sufficient Amounts of Viral E1" (2004). All Volumes (2001-2008). 86.