Hippenmeyer P J, Rankin A M, Reitz B A, McWilliams D R, Brightwell B B, Wolfe R A, Warren T G
G.D. Searle and Company, Research and Development, St. Louis, MO 63198, USA.
Mol Cell Endocrinol. 1995 Feb;107(2):155-64. doi: 10.1016/0303-7207(94)03439-z.
We have engineered GH3 cells with reporter genes under control of the growth hormone and prolactin promoters and measured protein production. The results indicate very low level production of reporter proteins from the cells regardless of the promoter used to drive expression. This was surprising in light of the observation that the cells still produced high levels of endogenous growth hormone and prolactin. Chinese hamster ovary (CHO) cells were engineered to express the Pit-1 transactivator. Transfection of reporter genes under control of the prolactin promoter demonstrated a clear enhancement of expression levels compared to the same promoter in parental CHO cells. Pit-1 expression is not sufficient, however, for high level, stable expression from the growth hormone promoter. These results indicate that the growth hormone and prolactin promoters are not sufficient for high level, stable expression even in normally permissive cells and suggest that Pit-1 alone is not sufficient for strong promoter activity from the integrated plasmids.
我们构建了在生长激素和催乳素启动子控制下带有报告基因的GH3细胞,并检测了蛋白质产生情况。结果表明,无论使用何种启动子来驱动表达,这些细胞产生的报告蛋白水平都非常低。鉴于观察到这些细胞仍能产生高水平的内源性生长激素和催乳素,这一结果令人惊讶。中国仓鼠卵巢(CHO)细胞被构建用于表达Pit-1反式激活因子。与亲代CHO细胞中相同启动子相比,在催乳素启动子控制下转染报告基因显示出表达水平明显增强。然而,Pit-1的表达对于生长激素启动子的高水平、稳定表达并不足够。这些结果表明,即使在通常允许的细胞中,生长激素和催乳素启动子也不足以实现高水平、稳定的表达,并且提示单独的Pit-1对于整合质粒的强启动子活性是不够的。