Chang B D, Roninson I B
Department of Genetics, University of Illinois at Chicago 60607-7170, USA.
Gene. 1996 Dec 12;183(1-2):137-42. doi: 10.1016/s0378-1119(96)00532-x.
We have developed lac repressor-regulated retroviral expression vectors that are induced by beta-galactosides upon stable transduction in mammalian cells. These vectors, derived from a Moloney-virus-based vector LNCX, contain an internal Rous sarcoma virus (RSV) or cytomegalovirus (CMV) promoter, coupled with 2 to 4 lac operator sequences and placed in anti orientation relative to the retroviral long terminal repeat (LTR). Three different vectors were tested in stably infected mass populations of mouse and human cells expressing the lac repressor, in parallel with the constitutively expressed LNCX vector. The highest expression levels from these vectors ranged from 1-4% to 25-33% of the LNCX level, and the induction by beta-galactosides ranged from 6-11-fold to 29-54-fold. These vectors should be suitable for studies requiring efficient gene transfer and regulated expression in mass populations of stably transduced mammalian cells.
我们开发了乳糖阻遏物调控的逆转录病毒表达载体，该载体在哺乳动物细胞中稳定转导后可被β-半乳糖苷诱导。这些载体衍生自基于莫洛尼病毒的载体LNCX，包含一个内部劳氏肉瘤病毒（RSV）或巨细胞病毒（CMV）启动子，与2至4个乳糖操纵序列相连，并以与逆转录病毒长末端重复序列（LTR）相反的方向放置。在稳定感染表达乳糖阻遏物的小鼠和人类细胞群体中，对三种不同的载体进行了测试，并与组成型表达的LNCX载体进行了平行比较。这些载体的最高表达水平为LNCX水平的1-4%至25-33%，β-半乳糖苷的诱导倍数为6-11倍至29-54倍。这些载体应适用于需要在稳定转导的哺乳动物细胞群体中进行高效基因转移和调控表达的研究。
