pBR322 contains glucocorticoid regulatory element DNA consensus sequences.
pBR322 contains glucocorticoid regulatory element DNA consensus sequences.
复制标题
pBR322 含有糖皮质激素调节元件 DNA 共有序列。
DOI：
10.1016/s0006-291x(87)80467-9
复制
发表时间：
1987
影响因子：
3.1
通讯作者：
Cidlowski,JA
中科院分区：
文献类型：
--
作者：
Tully,DB;Cidlowski,JA
对pBR322 DNA序列的计算机搜索发现了与已报道的糖皮质激素调节元件(GRE)DNA共有序列匹配的五个位点和三个相关位点。含有一个GRE位点的pBR322 DNA片段可与固定化的HeLa S3细胞糖皮质激素受体结合，并在竞争结合试验中竞争结合受体。相反，在这两种检测中，缺乏GRE位点的pBR322 DNA片段几乎检测不到与糖皮质激素受体的相互作用。这些结果证明了GRE共有序列在糖皮质激素受体与DNA相互作用中的重要性，并进一步确定了在糖皮质激素受体-DNA相互作用研究中将pBR322 DNA用作阴性对照时观察到高背景结合的原因。
A computer search of the pBR322 DNA sequence identified five sites matching reported glucocorticoid regulatory element (GRE) DNA consensus sequences and three related sites. A pBR322 DNA fragment containing one GRE site was shown to bind immobilized HeLa S3cell glucocorticoid receptor and to compete for receptor binding in a competitive binding assay. Conversely, a pBR322 DNA fragment devoid of GRE sites showed barely detectable interaction with glucocorticoid receptor in either of these assays. These results demonstrate the importance of GRE consensus sequences in glucocorticoid receptor interactions with DNA, and further identify a cause for high background binding observed when pBR322 DNA is used as a negative control in studies of glucocorticoid receptor-DNA interactions.