CHARACTERIZATION AND GROWTH FACTOR FOR MATURE CELLS

成熟细胞的特征和生长因子

• 批准号: 3488407
• 负责人: CHRISTOPHER S. HENNEY
• 金额: $ 5万
• 依托单位: IMMUNEX CORPORATION
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-05-01 至 1986-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3488407
• 关键词: B lymphocyte; Escherichia coli; chromatography; gel electrophoresis; gene expression; genetic library; genetic translation; growth factor; leukocyte activation /transformation; leukocyte disorder; messenger RNA; molecular cloning; mouse leukemia; neoplastic cell culture for noncancer research; tissue /cell culture; yeasts

The long-term (Phase II) objective of this proposal is the molecular characterization (cDNA cloning, expression) of murine and human genes coding for a growth factor for mature B-lymphocytes (m-BC-GF). Our approach will be to construct a cDNA library from sized, active, m-BC-GF mRNA and to isolate a full-length m-BC-GF cDNA by hybridization with an oligonucleotide probe deduced from protein sequence information derived from purified m-BC-GF protein. During Phase I of this proposal, m-BC-GF activity will be purified from supernates produced by a cloned murine lymphoma cell line (EL-4 lx cell line) recently developed in our laboratory. mRNA coding for m-BC-GF activity in vitro will also be prepared from the EL-4 lx cell line and size fractionated in preparation for cDNA cloning. Once murine m-BC-GF cDNAs are isolated, they will be expressed in appropriate yeast and E. coli vectors. Recombinant m-BC-GF will then be tested for biological activity in murine model systems of B-cell deficiency. Finally, murine cDNAs for m-BC-GF will be used as probes for identification by Southern blot hybridization of analogous human cDNAs. m-BC-GF could be of value in clinical restoration of antibody responsiveness in cases of viral, chemotherapeutic, or radiation induced immune suppression, and for the in vitro generation of human B-lymphocytes and/or hybridomas.

该提案的长期（II期）目标是分子