Zinn S A, Ebert K M, Mehta N D, Joshi J, Kilpatrick D L
Worcester Foundation for Experimental Biology, Shrewsburg, Massachusetts 01545.
J Biol Chem. 1991 Dec 15;266(35):23850-5.
The rat and mouse proenkephalin genes each contains two distinct promoters, one of which is utilized exclusively by spermatogenic cells. The germ cell-specific promoter lacks TATA sequences, is G+C rich, and contains multiple initiation sites. To investigate the nature of the cis-acting elements that determine selective transcription of the proenkephalin gene in male germ cells, two rat proenkephalin-chloramphenicol acetyltransferase fusion genes containing the two different promoter regions as well as 1.6 or 0.3 kilobases, respectively, of 5'-flanking sequence were expressed in transgenic mice. Multiple transgenic lines were developed which expressed the fusion genes in testis, brain, and heart but not in tissues that do not normally express the proenkephalin gene. Fusion gene transcripts in transgenic mouse testes were localized to those spermatogenic cell types that utilize the spermatogenic cell promoter and were selectively and accurately initiated from the multiple rat germ cell start sites. Transgenic mice thus provide a useful model for the localization and characterization of cis-acting elements mediating transcription of the proenkephalin gene from its germ cell-specific promoter.
大鼠和小鼠的前脑啡肽原基因各自包含两个不同的启动子,其中一个仅由生精细胞使用。生殖细胞特异性启动子缺乏TATA序列,富含G+C,并且包含多个起始位点。为了研究决定前脑啡肽原基因在雄性生殖细胞中选择性转录的顺式作用元件的性质,将两个分别包含两个不同启动子区域以及1.6或0.3千碱基5'侧翼序列的大鼠前脑啡肽原-氯霉素乙酰转移酶融合基因在转基因小鼠中表达。培育出了多个转基因品系,这些品系在睾丸、脑和心脏中表达融合基因,但在正常情况下不表达前脑啡肽原基因的组织中不表达。转基因小鼠睾丸中的融合基因转录本定位于那些使用生精细胞启动子的生精细胞类型,并从多个大鼠生殖细胞起始位点选择性且准确地起始。因此,转基因小鼠为定位和表征介导前脑啡肽原基因从其生殖细胞特异性启动子转录的顺式作用元件提供了一个有用的模型。