Isolation of Sparus auratus prolactin gene and activity of the cis-acting regulatory elements
Isolation of Sparus auratus prolactin gene and activity of the cis-acting regulatory elements
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DOI：
10.1016/s0016-6480(03)00214-4
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发表时间：
2003-10-15
影响因子：
2.7
通讯作者：
Valdivia, MM
中科院分区：
文献类型：
--
作者：
Astola, A;Ortiz, M;Valdivia, MM
使用 PCR 产生的催乳素 cDNA 片段作为探针分离海鲷催乳素 (sbPRL) 基因。分析的基因包含 3.5 kb 的 DNA，其中包含 5 个外显子，如之前针对其他鱼类 PRL 基因所述的那样。对 1.0 kb 近端启动子序列的分析揭示了共有 TATAA 盒、多达 7 个用于结合垂体特异性因子 Pit-1 的 (A/T)(3)NCAT 共有基序以及推定的 CREB 和 GATA 结合位点。用 sbPRL 启动子序列和海鲷 Pit-1 cDNA 表达质粒共转染的 CHO 培养细胞显示出连接的荧光素酶报告基因的表达。 5'-缺失突变体的瞬时表达实验揭示了 sbPRL 基因上至少三个调控区域，其中两个对转录具有刺激作用，一个具有明显的抑制作用。从比较的角度来看，这项对黑鲷 PRL 基因的研究与之前发表的罗非鱼和虹鳟鱼 PRL 基因的研究具有很好的相关性。报告的分子数据将有助于硬骨鱼 GH/PRL 基因家族基因调控的比较分析。 (C) 2003 年爱思唯尔科学（美国）。版权所有。
A sea bream prolactin (sbPRL) gene was isolated using a prolactin cDNA fragment, generated by PCR as a probe. The gene analyzed comprises 3.5 kb of DNA containing five exons as described previously for other fish PRL genes. Analysis of 1.0 kb of the proximal promoter sequence reveals a consensus TATAA box, up to seven (A/T)(3)NCAT consensus motifs for binding of the pituitary-specific factor Pit-1 and putative CREB and GATA binding sites. CHO culture cells co-transfected with a sbPRL promoter sequence and a sea bream Pit-1 cDNA expression plasmid showed expression of a linked luciferase reporter gene. Transient expression experiments with 5'-delection mutants reveals at least three regulatory regions on the sbPRL gene, two with a stimulatory effect on transcription and one with apparent inhibitory effect. From a comparative point of view, this study of PRL gene in Sparus auratus, correlates well with those previously published on tilapia and rainbow trout. The molecular data reported will be useful for comparative analysis of gene regulation in the GH/PRL gene family in teleosts. (C) 2003 Elsevier Science (USA). All rights reserved.