COMPARISON OF SEVERAL PROMOTERS AND POLYADENYLATION SIGNALS FOR USE IN HETEROLOGOUS GENE-EXPRESSION IN CULTURED DROSOPHILA CELLS
COMPARISON OF SEVERAL PROMOTERS AND POLYADENYLATION SIGNALS FOR USE IN HETEROLOGOUS GENE-EXPRESSION IN CULTURED DROSOPHILA CELLS
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DOI：
10.1093/nar/19.18.5037
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发表时间：
1991-09-25
影响因子：
14.9
通讯作者：
IVEYHOYLE, M
中科院分区：
文献类型：
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作者：
ANGELICHIO, ML;BECK, JA;IVEYHOYLE, M
我们直接比较了四个启动子和三个聚腺苷酸化 (poly(A)) 信号在稳定转染的果蝇 S2 细胞中指导异源基因表达的能力。 我们将两种组成型果蝇启动子（肌动蛋白 5C 远端启动子和 α-1-微管蛋白启动子）与严格调控的果蝇金属硫蛋白 (Mtn) 启动子和家蚕丝蛋白启动子进行了比较。 我们发现肌动蛋白 5c 和诱导的 Mtn 启动子在该系统中产生相当高水平的 RNA 和蛋白质。 α-1-微管蛋白启动子产生的水平大约低四倍，丝心蛋白启动子在 S2 细胞中没有显示出可检测到的活性。 有趣的是，组成型肌动蛋白 5C 和 α-1-微管蛋白启动子表达的基因的拷贝数始终比诱导型 Mtn 启动子或无活性丝心蛋白启动子表达的基因低三到四倍。 哺乳动物 (SV40) 和果蝇 (Mtn) 来源的 Poly(A) 信号可有效指导 S2 细胞中稳定的 RNA 合成，并且与哺乳动物细胞一样，SV40 晚期 Poly(A) 信号比 SV40 早期 Poly(A) 信号更有效。 因此，多腺苷酸化过程似乎在哺乳动物和果蝇细胞之间是保守的。
We have directly compared the ability of four promoters and three polyadenylation (poly(A)) signals to direct heterologous gene expression in stably transfected Drosophila melanogaster S2 cells. We compared two constitutive Drosophila promoters, the actin 5C distal promoter and the alpha-1-tubulin promoter, with the tightly regulated Drosophila metallothionein (Mtn) promoter and the Bombyx mori fibroin promoter. We find that the actin 5c and induced Mtn promoters generate comparable high levels of RNA and protein in this system. The alpha-1-tubulin promoter generates about four-fold lower levels, and the fibroin promoter shows no detectable activity in S2 cells. Interestingly, genes expressed from the constitutive actin 5C and alpha-1-tubulin promoters are consistently present at three- to four-fold lower copy numbers than genes expressed from the inducible Mtn promoter or the inactive fibroin promoter. Poly(A) signals of both mammalian (SV40) and Drosophila (Mtn) origin efficiently directed stable RNA synthesis in S2 cells, and, as in mammalian cells, the SV40 late poly(A) signal was more efficient than the SV40 early poly(A) signal. Thus the process of polyadenylation appears to be conserved between mammalian and Drosophila cells.