Core--Vector

载体核心将产生基于单纯疱疹病毒（HSV）的扩增子载体、腺病毒载体和逆转录病毒载体。这些将用于项目1中的混合神经元/神经胶质细胞培养物和纯星形胶质细胞培养物，以及项目2中的体内缺血研究。将产生HSV和腺病毒载体，其表达CuZn-超氧化物歧化酶（SOD 1）、谷胱甘肽过氧化物酶（前两者单独或组合）、Bcl-2和Hsp 70的基因。所有的HSV和腺病毒载体都是"双顺反子"的，既表达所讨论的基因，也表达报告基因。对于每个实验载体，同源对照载体将含有所讨论的相同基因，在其中心插入终止密码子，确保不表达。对于腺病毒载体将产生类似的对照。将生产表达SOD 1、Mn-SOD（SOD 2）、Bcl-2和Hsp 70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形式。HSP70和Bcl-2的逆转录病毒载体已经构建，它们将在载体核心中产生。此外，将构建并随后产生表达SOD1和SOD2（分别）的新载体。载体核心的目的是确保为各个群体不断提供载体，确保对这些载体生产的质量控制，并在各个实验室使用载体时解决出现的问题。

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.