Clegg D O
Neuroscience Research Institute, University of California, Santa Barbara 93106.
Gene. 1993 Aug 25;130(2):291-6. doi: 10.1016/0378-1119(93)90434-5.
A mammalian expression vector that directs expression of murine beta-nerve growth factor (beta-NGF) from a murine sarcoma virus long terminal repeat (LTR) promoter element was constructed and characterized. The vector, designated pLTRSNGF, was stably transfected into murine L-cells, and beta-NGF mRNA and protein levels were quantified and compared to endogenous levels in control L-cells. Transfection of pLTRSNGF resulted in an approximate doubling of both beta-NGF mRNA and mature beta-NGF protein secreted into the media. Transfection of pLTRSNGF into rat PC 12 cells resulted in colonies of autocrine-differentiating cells that extended dense networks of neurites in the absence of added NGF, indicating that the beta-NGF produced from the vector is biologically active.
构建并表征了一种哺乳动物表达载体，该载体从鼠肉瘤病毒长末端重复序列（LTR）启动子元件指导鼠β-神经生长因子（β-NGF）的表达。将该载体命名为pLTRSNGF，稳定转染至鼠L细胞中，对β-NGF mRNA和蛋白质水平进行定量，并与对照L细胞中的内源性水平进行比较。pLTRSNGF的转染导致分泌到培养基中的β-NGF mRNA和成熟β-NGF蛋白质均增加约一倍。将pLTRSNGF转染至大鼠PC 12细胞中，产生了自分泌分化细胞集落，这些细胞在未添加NGF的情况下延伸出密集的神经突网络，表明该载体产生的β-NGF具有生物活性。
