Musti A M, Ursini V M, Avvedimento E V, Zimarino V, Di Lauro R
Centro di Endocrinologia ed Oncologia sperimentale del CNR, II Facoltà di Medicina, Napoli, Italy.
Nucleic Acids Res. 1987 Oct 26;15(20):8149-66. doi: 10.1093/nar/15.20.8149.
We have fused a 900 base pair long DNA segment containing the transcriptional start site of the rat thyroglobulin (Tg) gene to the bacterial gene for chloramphenicol acetyltransferase (cat). The fusion gene has been introduced into three different cell lines derived from the rat thyroid gland and into a rat liver cell line. Expression of the fusion gene was detected only in the one thyroid cell line that is able to express the endogenous Tg gene. The minimum DNA sequence required for the cell type specific expression was determined by deletion analysis; it extends 170 nucleotides upstream of the transcription initiation site. The Tg promoter contains a readily detectable binding sites for a factor present in salt extracts of thyroid cell nuclei. This binding site is not recognized by the nuclear extracts of any other cell type that we have tested, suggesting that it may help mediate the cell type specific expression of the Tg gene.
我们将一段包含大鼠甲状腺球蛋白(Tg)基因转录起始位点的900个碱基对长的DNA片段与氯霉素乙酰转移酶(cat)的细菌基因融合。该融合基因已被导入源自大鼠甲状腺的三种不同细胞系以及一种大鼠肝细胞系。仅在能够表达内源性Tg基因的一种甲状腺细胞系中检测到了融合基因的表达。通过缺失分析确定了细胞类型特异性表达所需的最小DNA序列;它在转录起始位点上游延伸170个核苷酸。Tg启动子含有一个易于检测到的甲状腺细胞核盐提取物中存在的因子的结合位点。我们测试的任何其他细胞类型的核提取物都无法识别该结合位点,这表明它可能有助于介导Tg基因的细胞类型特异性表达。