Somasekhar M B, Gorski J
Department of Biochemistry, College of Agriculture and Life Sciences, University of Wisconsin-Madison 53706.
Gene. 1988 Sep 15;69(1):13-21. doi: 10.1016/0378-1119(88)90373-3.
The 5'-flanking region of the rat prolactin gene contains two DNase I-hypersensitive (HS) sites. We used gene transfer experiments to determine the nucleotide (nt) sequences within and around these two HS sites that may contain the information necessary for regulation of prolactin gene expression by estrogens and glucocorticoids. A chimeric gene construct (pPRL.CAT) was prepared by covalently linking the sequence of the rat prolactin gene to the bacterial chloramphenicol acetyltransferase-coding gene, cat. Rat GH3 cells were transfected with pPRL.CAT and six mutants that possess deletions within and around the two HS sites. Incubation of such transfectants with estrogen or dexamethasone indicated the existence of two functionally important elements within the 5'-flanking region of the rat prolactin gene. The element required for estrogen up-regulation of the prolactin gene is located between nt residues -1530 through -1950. The glucocorticoid down-regulatory element is located between nt -200 and +75.
大鼠催乳素基因的5'侧翼区含有两个对DNA酶I敏感的（HS）位点。我们利用基因转移实验来确定这两个HS位点及其周围可能包含雌激素和糖皮质激素调节催乳素基因表达所需信息的核苷酸（nt）序列。通过将大鼠催乳素基因的序列与细菌氯霉素乙酰转移酶编码基因cat共价连接，制备了一个嵌合基因构建体（pPRL.CAT）。用pPRL.CAT和六个在两个HS位点及其周围具有缺失的突变体转染大鼠GH3细胞。用雌激素或地塞米松培养这些转染体表明，在大鼠催乳素基因的5'侧翼区内存在两个功能重要的元件。催乳素基因雌激素上调所需的元件位于nt残基-1530至-1950之间。糖皮质激素下调元件位于nt -200至+75之间。
