Noteborn M H, Verschueren C A, Zantema A, Koch G, van der Eb A J
Laboratory for Molecular Carcinogenesis, Sylvius Laboratory, Leiden University, The Netherlands.
Gene. 1994 Dec 15;150(2):313-8. doi: 10.1016/0378-1119(94)90444-8.
The single promoter region in the cloned genome [Noteborn et al., J. Virol. 65 (1991) 3131-3139] of chicken anemia virus (CAV) in chicken T-cells was analysed via CAT assays. A unique region containing four or five near-perfect direct repeats (DR) of 21 bp with one 12-bp insert was proven to be the main transcription-activation element, with enhancer-like characteristics. PCR studies revealed that CAV isolates from across the world all contained this promoter sequence. Electrophoretic mobility-shift assays (EMSA) showed that individual DR units, as well as the 12-bp insert, can bind to nuclear factors of chicken T-cells. Competition assays revealed that the DR units bound to factors other than the 12-bp insert. A synthetic oligodeoxyribonucleotide containing an SP1-box (5'-GGGCGG) could compete with factors binding to the 12-bp insert. Purified human SP1 was shown to have very strong affinity for the 12-bp insert.
通过氯霉素乙酰转移酶（CAT）分析，对鸡贫血病毒（CAV）在鸡T细胞中的克隆基因组[诺特博恩等人，《病毒学杂志》65（1991）3131 - 3139]中的单个启动子区域进行了分析。一个独特的区域被证明是主要的转录激活元件，具有增强子样特征，该区域包含四个或五个21 bp的近乎完美的直接重复序列（DR），中间有一个12 bp的插入序列。聚合酶链反应（PCR）研究表明，来自世界各地的CAV分离株都含有这个启动子序列。电泳迁移率变动分析（EMSA）显示，单个DR单元以及12 bp的插入序列都能与鸡T细胞的核因子结合。竞争分析表明，DR单元与12 bp插入序列以外的因子结合。一个含有SP1框（5'-GGGCGG）的合成寡脱氧核糖核苷酸可以与结合到12 bp插入序列的因子竞争。纯化的人SP1对12 bp插入序列显示出很强的亲和力。
