Krimpenfort P, de Jong R, Uematsu Y, Dembic Z, Ryser S, von Boehmer H, Steinmetz M, Berns A
Division of Molecular Genetics, the Netherlands Cancer Institute, Amsterdam.
EMBO J. 1988 Mar;7(3):745-50. doi: 10.1002/j.1460-2075.1988.tb02871.x.
To characterize cis-acting elements controlling the expression of T cell receptor beta-chains we generated a number of transgenic mouse lines harboring a rearranged T cell receptor beta-chain with different extensions of 5' and 3' flanking sequences. Transcriptional analysis of transgenic mice carrying these clones showed that sequences located downstream of the polyadenylation signal of the C beta 2 region are indispensable for expression in transgenic mice. The sequences conferring enhancer activity in this fragment were further defined by transient CAT assays. Strong enhancer activity was found to reside in a 550 bp fragment located 5 kb downstream from C beta 2. The nucleotide sequence of this fragment revealed a number of oligonucleotide motifs characteristic for enhancer elements.
为了鉴定控制T细胞受体β链表达的顺式作用元件,我们构建了许多转基因小鼠品系,这些品系携带重排的T细胞受体β链,并带有不同长度的5'和3'侧翼序列。对携带这些克隆的转基因小鼠进行转录分析表明,位于Cβ2区域聚腺苷酸化信号下游的序列对于在转基因小鼠中的表达是必不可少的。通过瞬时CAT分析进一步确定了该片段中赋予增强子活性的序列。发现强增强子活性存在于位于Cβ2下游5 kb处的一个550 bp片段中。该片段的核苷酸序列揭示了一些增强子元件特有的寡核苷酸基序。