INTRODUCTION OF THE HIV TAT GENE INTO LYMPHOID CELLS

将 HIV TAT 基因引入淋巴细胞

• 批准号: 3896376
• 负责人: J A LAUTENBERGER
• 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/3896376
• 关键词: AIDS; Retroviridae; gene expression; genetic manipulation; genetic regulatory element; genetic transcription; human immunodeficiency virus; lymphocyte; neoplastic cell culture for noncancer research; plasmids; tissue /cell culture; transfection; transposon /insertion element; virus genetics; virus protein; virus replication

In order to study the effect of the human immunodeficiency virus (HIV) tat gene on the expression of cellular genes, we have constructed lymphoid lines that expressed the tat gene. This was done by placing the tat gene and an adjacent HIV long terminal repeat (LTR) into the retroviral vector pGV1. The recombinant plasmid was transfected into psi2 cells to produce an ecotropic viral stock that was used to infect psiAM cells. Colonies of G418-resistant psiAM cells were isolated and assayed for the production of virus. Supernatants of those colonies with the highest virus production were used to infect cells of the human T-cell lymphoid line, H9. G418-resistant cell lines derived from H9 were found to contain tat activity as assayed by fusion with HeLa cells containing LTR and CAT sequences. Furthermore, they contained an LTR-tat message of the predicted size.

为了研究人类免疫缺陷病毒（HIV）的影响 基因对细胞基因表达的影响，我们构建了淋巴 表达 tat 基因的品系。这是通过将 tat 基因置于 和一个相邻的 HIV 长末端重复序列 (LTR) 进入逆转录病毒载体 pGV1。将重组质粒转染至psi2细胞中，产生 用于感染 psiAM 细胞的亲嗜性病毒原种。菌落 分离并分析 G418 抗性 psiAM 细胞的生产情况 病毒。 使用病毒产量最高的菌落的上清液 感染人类 T 细胞淋巴系 H9 的细胞。 G418抗性细胞 经测定，发现源自 H9 的品系含有 tat 活性 与含有 LTR 和 CAT 序列的 HeLa 细胞融合。此外，他们 包含预测大小的 LTR-tat 消息。