Core--Vector

载体核心将生产基于单纯疱疹病毒(HSV)的扩增载体、腺病毒载体和逆转录病毒载体。这些将在项目1中用于混合神经元/神经胶质细胞培养和纯星形胶质细胞培养，以及在项目2中用于活体缺血研究。单纯疱疹病毒载体和腺病毒载体将分别表达铜锌超氧化物歧化酶(SOD1)和谷胱甘肽过氧化物酶(前两者)的基因，或者单独表达，或者联合表达，即Bcl-2和Hsp70。所有的单纯疱疹病毒和腺病毒载体都将是“双顺反子”的，既表达有问题的基因，也表达报告基因。对于每个实验载体，同源对照载体将包含相同的基因，并在其中心插入一个终止密码子，以确保不表达。对腺病毒载体也将产生类似的对照。将产生表达SOD1、Mn-SOD(SOD2)、Bcl2和Hsp70的逆转录病毒载体。对照载体将表达报告基因β-半乳糖苷酶或目的基因的终止密码子版本。HSP70和Bcl2逆转录病毒载体已经构建，并将在载体核心产生。此外，还将构建和生产分别表达SOD1和SOD2的新载体。病媒核心的目的将是确保为不同群体持续供应病媒，确保在生产这种病媒的过程中进行质量控制，并在个别实验室使用病媒时解决出现的问题。

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.