EXPRESSION OF HTLV-III ENVELOPE GENE IN EUKARYOTIC (SV40)-BASED EXPRESSION VECTOR

HTLV-III 包膜基因在真核 (SV40) 表达载体中的表达

• 批准号: 3963579
• 负责人: T S PAPAS
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3963579
• 关键词: cell bank /registry; embryo /fetus; eukaryote; gene expression; genetic manipulation; genetic recombination; human immunodeficiency virus 1; immunochemistry; metallothionein; simian virus 40; virus envelope; virus genetics

Several constructs using the power of recombinant DNA technology were employed to express in eukaryotic vector systems the envelope genes encoded by the HTLV-III virus. One type of construct incoporates a portion of the HTLV-III envelope genes fused to an SV40 promoter region, complete with splice and polyadenylation signals. A second type of expression vector construct fused the HTLV-III viral envelope gene with the SV40 T antigen amino terminal region, which can elicit proteins recognized by specific antibodies directly against T antigen. Both types of constructs, after contransfection into TK- cells along with the thymidine kinase gene, gave rise to selective transformants that enabled permanent lines to be established. In particular, 8 out of 10 such cell lines were found to be expressing HTLV-III envelope specific mRNA. Four of these cell lines were expressing high levels of message, while the other four were moderately expressive. Recombinant vectors containing metallothionein promoter inducible HTLV-III envelope genes were recently constructed conatining SV40 T antigen splice and polyadenylation signals and are being introduced into embryonic mice for transgenic expression studies.

利用重组DNA技术的力量构建了几种构建体， 用于在真核载体系统中表达编码的包膜基因 HTLV-III病毒感染。 一种类型的构造包括一部分的 HTLV-III包膜基因融合到SV 40启动子区，完整的 剪接和多聚腺苷酸化信号。 第二类表达载体 将HTLV-III病毒包膜基因与SV 40 T抗原融合的构建体 氨基末端区域，其可以引发特异性 直接对抗T抗原的抗体。 这两种类型的结构， 与胸苷激酶基因沿着共转染TK-细胞， 产生选择性的转化体，使永久株系能够 确立了习 特别地，发现10个这样的细胞系中有8个是 表达HTLV-III包膜特异性mRNA。 这些细胞系中有四种是 表达高水平的信息，而其他四个是中等 富有表现力。 含有金属硫蛋白启动子的重组载体 最近构建了含有SV 40的可诱导HTLV-III包膜基因 T抗原剪接和多聚腺苷酸化信号，并被引入 用于转基因表达研究的胚胎小鼠。