The Vector core will produce herpes simplex virus (HSV)-based amplicon vectors, adenoviral vectors, and retroviral vectors. These will be used in Project 1 in mixed neuronal/glial cultures and pure astrocyte cultures, as well as in Project 2, for in vivo ischemia studies. HSV and adenoviral vectors will be generated expressing the genes for CuZn- superoxide dismutase (SOD1), glutathione peroxidase (the prior two either individually, or in combination, Bcl-2, and Hsp70. All of the HSV and adenoviral vectors will be """"""""bicistronic"""""""", expressing both the gene in question, as well as a reporter gene. For each experimental vector, the cognate control vector will contain the same gene in question with a stop codon inserted in its center, insuring non-expression. Similar controls will be generated for the adenoviral vectors. Retroviral vectors to express SOD1, Mn-SOD (SOD2), Bcl-2 and Hsp70 will be produced. Control vector will express either the reporter gene beta-galactosidase or a stop codon version of the gene of interest. The retroviral vectors for HSP70 and Bcl-2 have been constructed and they will be produced in the vector core. In addition new vectors to express SOD1 and SOD2 (separately) will be constructed and then produced. The purpose of the Vector Core will be to ensure a constant supply of vectors for the various groups, to insure quality control in the production of such vectors, and to troubleshoot problems, as they arise, in individual laboratories in the use of vectors.
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