COBRE: UNV MED SCH: CORE B: MOLECULAR EXPRESSION & TRANGENICS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。分子表达和转基因核心（核心B）将具有三个主要功能：1）提供腺病毒基因转移载体和蛋白转导试剂; 2）通过RT-PCR和蛋白质印迹法评估所有项目的mRNA和蛋白质表达; 3）获得转基因小鼠并维持转基因小鼠品系。William T.格霍弗将指挥核心。他将与研究助理教授Cherie Singer博士和两名高级研究员密切合作，组织核心实验室的日常活动。总监将在每月进度报告中与项目PI协商，并解决基因转移和蛋白转导载体的生产或质量问题。研究助理教授Cherie Singer博士将设计所有要求的质粒和病毒构建体，并将监督研究助理Shanti Rawat，他将担任载体生产的首席技术员。Rawat夫人将克隆感兴趣的基因，生产用于蛋白质表达和质粒繁殖的细菌细胞，并构建腺病毒载体。她将由第二名全职技术员Mariam Ba协助，她将协助繁殖细菌和病毒载体以及病毒包装细胞。Ba女士还将协助项目PI通过RT-PCR和蛋白质印迹法评估培养的平滑肌细胞和组织中的转基因表达。她还接受了基本肌肉力学和原位基因传递组织电穿孔方法的培训。核心设备（通风柜、培养箱、水浴、离心机）也将由核心B工作人员维护。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Molecular Expression and Transgenic Core (Core B) will have three major functions: 1) To provide adenoviral gene transfer vectors and protein transduction reagents; 2) To assess mRNA and protein expression for all projects by RT-PCR and western blotting and 3) To procure transgenic mice and maintain transgenic mouse lines. Dr. William T. Gerthoffer will direct the core. He will work closely with Dr. Cherie Singer, Research Assistant Professor and two senior Research Associates to organize daily activities of the Core lab. The Director will consult with Project PIs during monthly progress reports and will solve problems that arise with production or quality of the gene transfer and protein transduction vectors. Dr. Cherie Singer, Research Assistant Professor will design all requested plasmid and virus constructs and will supervise Shanti Rawat, Research Associate who will serve as the chief technician for vector production. Mrs. Rawat will clone genes of interest, produce bacterial ceils for protein expression and propagation of plasmids and build adenovirus vectors. She will be assisted by a second full-time technician, Mariam Ba, who will assist in propagating bacterial and viral vectors and viral packaging cells. Ms Ba will also assist project PIs in assessing transgene expression in cultured smooth muscle cells and tissues by RT-PCR and western blotting. She is also trained in basic muscle mechanics and tissue electroporation methods for gene delivery in situ. Core equipment (hoods, incubator, water bath, centrifuges)will also be maintained by the Core B staff.