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小鼠β4-半乳糖基转移酶在雄性生殖细胞中的表达。一个包含两个环磷酸腺苷反应元件(CRE)样元件的796个碱基对的基因组区域,介导转基因小鼠中雄性生殖细胞特异性表达。

Male germ cell expression of murine beta 4-galactosyltransferase. A 796-base pair genomic region, containing two cAMP-responsive element (CRE)-like elements, mediates male germ cell-specific expression in transgenic mice.

作者信息

Shaper N L, Harduin-Lepers A, Shaper J H

机构信息

Cell Structure and Function Laboratory, Johns Hopkins Oncology Center, Baltimore, Maryland.

出版信息

J Biol Chem. 1994 Oct 7;269(40):25165-71.

PMID:7929205
Abstract

In murine somatic cells, transcription of the single gene encoding beta 4-galactosyltransferase results in two transcripts of 4.1 and 3.9 kilobases (kb), as a consequence of the use of two transcriptional start sites that are located on exon one separated by 200 base pairs (bp). In early male germ cell development, spermatogonia use only the 4.1-kb start site to yield a transcript that is identical to its somatic cell counterpart. As these cells enter meiosis, there is a switch from the use of this somatic cell start site to the exclusive use, beginning in pachytene spermatocytes, of a male germ cell-specific start site. Germ cell-specific transcripts are distinguished from their somatic counterparts by an additional approximately 560 nucleotides of 5'-untranslated sequence that is located immediately upstream and contiguous with the transcriptional start site defined for the 4.1-kb mRNA (Harduin-Lepers, A., Shaper, N.L., Mahoney, J.A., and Shaper, J.H. (1992) Glycobiology 2, 361-368). This observation predicts the use of a different upstream male germ cell-specific promoter. In this study we show that a 796-bp fragment containing 543 bp of genomic sequence upstream of the germ cell specific transcriptional start site and 253 bp of flanking downstream sequence, directs expression of the reporter gene, beta-galactosidase, exclusively to the pachytene spermatocytes and round spermatids of transgenic mice. This pattern of cell type-specific expression of the transgene is comparable with that of the endogenous beta 4-galactosyltransferase gene.

摘要

在鼠类体细胞中,由于使用了位于外显子1上、相隔200个碱基对(bp)的两个转录起始位点,编码β4-半乳糖基转移酶的单基因转录产生了4.1千碱基(kb)和3.9 kb的两种转录本。在雄性生殖细胞早期发育过程中,精原细胞仅使用4.1-kb的起始位点产生一种与其体细胞对应物相同的转录本。当这些细胞进入减数分裂时,起始位点的使用发生了转变,从体细胞起始位点的使用转变为从粗线期精母细胞开始专一使用雄性生殖细胞特异性起始位点。生殖细胞特异性转录本与其体细胞对应物的区别在于,其5'-非翻译序列额外增加了约560个核苷酸,该序列紧邻为4.1-kb mRNA定义的转录起始位点且位于其上游(哈迪因-勒佩斯,A.,沙珀,N.L.,马奥尼,J.A.,和沙珀,J.H.(1992年)《糖生物学》2,361 - 368)。这一观察结果预示着使用了一个不同的上游雄性生殖细胞特异性启动子。在本研究中,我们表明一个796-bp的片段(包含生殖细胞特异性转录起始位点上游543 bp的基因组序列和侧翼下游253 bp的序列)将报告基因β-半乳糖苷酶的表达专一性地导向转基因小鼠的粗线期精母细胞和圆形精子细胞。转基因这种细胞类型特异性表达模式与内源性β4-半乳糖基转移酶基因的模式相当。

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