Staudt L M, Clerc R G, Singh H, LeBowitz J H, Sharp P A, Baltimore D
Whitehead Institute for Biomedical Research, Cambridge, MA 02142.
Science. 1988 Jul 29;241(4865):577-80. doi: 10.1126/science.3399892.
An octamer DNA sequence plays a critical role in directing transcription of immunoglobulin genes in B lymphocytes. A new technique of direct binding of radioactive DNA was used to screen a complementary DNA expression library from the BJAB cell line in lambda gt11 phage to derive molecular cDNA clones representing a putative B lymphocyte-specific octamer binding protein. The plaques were screened with DNA containing four copies of the octamer sequence and positive phage recombinants were identified. The fusion protein produced on inducing a lysogen of one phage bound to a monomeric octamer probe. The cDNA insert from this phage hybridized to messenger RNA found in B lymphocytes, but not in most other cells. Thus, this cDNA derives from a gene (oct-2) that specifies an octamer binding protein expressed preferentially in B lymphocytes, proving that, for at least one gene, a cell-specific transcription factor exists and its amount is controlled through messenger RNA availability.
一个八聚体DNA序列在指导B淋巴细胞中免疫球蛋白基因的转录方面起着关键作用。一种放射性DNA直接结合的新技术被用于筛选来自λgt11噬菌体中BJAB细胞系的互补DNA表达文库,以获得代表假定的B淋巴细胞特异性八聚体结合蛋白的分子cDNA克隆。用含有四个八聚体序列拷贝的DNA筛选噬菌斑,并鉴定出阳性噬菌体重组体。诱导一种噬菌体的溶原菌产生的融合蛋白与单体八聚体探针结合。来自该噬菌体的cDNA插入片段与B淋巴细胞中发现的信使RNA杂交,但在大多数其他细胞中未杂交。因此,该cDNA来源于一个基因(oct-2),该基因指定了一种在B淋巴细胞中优先表达的八聚体结合蛋白,证明了至少对于一个基因而言,存在细胞特异性转录因子,并且其数量通过信使RNA的可获得性受到控制。