Xiong F, Chin R A, Hew C L
Research Institute, Hospital of Sick Children, Toronto, Ontario, Canada.
Mol Mar Biol Biotechnol. 1992 Apr;1(2):155-64.
A full-length chinook salmon (Oncorhynchus tschawytscha) prolactin (PRL) gene, the first genomic clone of a teleost prolactin, was isolated and fully sequenced. The chinook PRL genomic sequence spans 6.4 Kb, including 2.4 Kb of 5' flanking sequence, 3.0 Kb representing the five exons and four introns of the complete PRL gene, and 0.9 Kb of 3' flanking sequence. The transcriptional start site of the PRL gene was mapped through the agreement of both primer extension and S1 nuclease protection assay. The 5' flanking region of the PRL gene was searched for potential cis-acting elements based on the consensus binding site of trans-acting factor Pit-1, known to be involved in PRL gene expression in mammals. Functional analysis of PRL promoter by the transient transfection of several PRL promoter/CAT chimeric plasmids into rainbow trout pituitary cells suggests a functional PRL promoter whose cell-specific activity is most likely governed by both positive and negative mechanisms.
一条完整的奇努克鲑(Oncorhynchus tschawytscha)催乳素(PRL)基因,这是硬骨鱼催乳素的首个基因组克隆,被分离并进行了全序列测定。奇努克鲑PRL基因组序列跨度为6.4千碱基对(Kb),包括2.4 Kb的5'侧翼序列、代表完整PRL基因的五个外显子和四个内含子的3.0 Kb以及0.9 Kb的3'侧翼序列。通过引物延伸和S1核酸酶保护试验的一致性确定了PRL基因的转录起始位点。基于已知参与哺乳动物PRL基因表达的反式作用因子Pit-1的共有结合位点,在PRL基因的5'侧翼区域寻找潜在的顺式作用元件。通过将几种PRL启动子/CAT嵌合质粒瞬时转染到虹鳟垂体细胞中对PRL启动子进行功能分析,结果表明存在一个功能性PRL启动子,其细胞特异性活性很可能受正、负两种机制调控。