CHARACTERIZATION OF THE CELLULAR LOCALIZATION OF BASE

基地细胞定位的表征

• 批准号: 7610323
• 负责人: KRISTI A EGLAND
• 金额: $ 0.23万
• 依托单位: UNIVERSITY OF SOUTH DAKOTA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7610323
• 关键词: Amino Acid Sequence; Amino Acids; Breast Cancer Cell; Cancer cell line; Cell Extracts; Cells; Computer Retrieval of Information on Scientific Projects Database; Culture Media; Evaluation; Family; Funding; Genes; Grant; Hour; Human; Immunoblot Analysis; In Vitro; Institution; Libraries; Membrane Proteins; Monoclonal Antibodies; N-terminal; Normal tissue morphology; Oncogenes; Open Reading Frames; Peptide Signal Sequences; Plasmids; Proteins; Research; Research Personnel; Resources; Salivary Glands; Source; System; Tumor Markers; United States National Institutes of Health; Western Blotting; disorder risk; latherin; malignant breast neoplasm; outcome forecast; response; vector

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. Tumor markers provide information for assessing disease risk, prognosis, metastatic potential and evaluation of response to therapy. Previously, a new breast cancer gene designated BASE (breast cancer and salivary gland expression) has been identified by generating a library that was enriched for genes encoding secreted and membrane proteins. Of 62 normal tissues analyzed, BASE was only expressed in salivary gland. BASE was also expressed in human breast cancers. The deduced amino acid sequence of BASE predicts that it is secreted. BASE is predicted to have a 20 amino acid N-terminal signal sequence, and has homology to other secreted proteins, such as Latherin and the PLUNC family. To further characterize BASE, we need to verify its cellular localization. As a first step, an in vitro expression system using 293T cells and the ZR-75-1 breast cancer cell line will be developed. A plasmid will be constructed containing the BASE ORF cloned 5' of a Myc tag using the vector pcDNA3.1-Myc/His. This expression construct will be transfected into 293T and ZR-76-1 cells. After 48 hours, the culture medium and total cell extracts will be examined for the presence of BASE-Myc by Western immunoblot analysis using a monoclonal antibody against the Myc tag.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 肿瘤标记提供了评估疾病风险，预后，转移性潜力和对治疗反应的评估的信息。 Previously, a new breast cancer gene designated BASE (breast cancer and salivary gland expression) has been identified by generating a library that was enriched for genes encoding secreted and membrane proteins. 在分析的62个正常组织中，仅在唾液腺中表达碱。 碱还在人类乳腺癌中表达。 推导的碱酸序列预测其分泌。 BASE is predicted to have a 20 amino acid N-terminal signal sequence, and has homology to other secreted proteins, such as Latherin and the PLUNC family. 为了进一步表征碱，我们需要验证其细胞定位。 作为第一步，将开发使用293T细胞和ZR-75-1乳腺癌细胞系的体外表达系统。 将使用vector pcDNA3.1-myc/His构建质粒，其中构建包含MYC标签的基本ORF 5'。 该表达构建体将转染到293T和ZR-76-1细胞中。 After 48 hours, the culture medium and total cell extracts will be examined for the presence of BASE-Myc by Western immunoblot analysis using a monoclonal antibody against the Myc tag.