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小鼠血管紧张素II 2型受体基因的表达、基因组结构及转录

Expression, genomic organization, and transcription of the mouse angiotensin II type 2 receptor gene.

作者信息

Ichiki T, Inagami T

机构信息

Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tenn 37232, USA.

出版信息

Circ Res. 1995 May;76(5):693-700. doi: 10.1161/01.res.76.5.693.

Abstract

Although the rat angiotensin II type 2 receptor (AT2) was cloned and shown to be a member of the seven transmembrane domain-type receptor family, its signaling mechanism and biological roles have not been established. To acquire additional information on the structure and functions of AT2 genomic DNA, we cloned the mouse AT2 gene and examined its expression, transcription, and genomic organization. The amino acid sequence of the mouse AT2 cDNA showed a 98.5% sequence identity with the rat AT2. In mouse fetus, mRNA of the AT2 was highly expressed in the eviscerated carcass and brain. This expression decreased rapidly after birth. In 10-week-old mice, mRNA of the AT2 could be detected in the brain by Northern blot analysis. However, reverse transcription-polymerase chain reaction showed that mRNA of the AT2 was expressed in all organs examined, indicating that the AT2 is expressed at a low level in other organs. Southern blot analysis of the genomic DNA of the mouse liver digested with BamHI, EcoRI, and HindIII resulted in single bands, indicating that the AT2 gene probably exists at a single locus in the mouse genome. The nucleotide sequence of the AT2 gene (4.5 kb of the EcoRI fragment) revealed the presence of three exons. An entire coding sequence was included in the third exon. Primer extension experiments showed the presence of two transcription initiation sites in the mouse AT2 gene. A DNA segment of about 1.5 kb of the promoter region (-1497 to +56 bp) of the mouse AT2 gene was fused to a luciferase reporter gene.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

尽管大鼠血管紧张素II 2型受体(AT2)已被克隆,并被证明是七跨膜结构域型受体家族的成员,但其信号传导机制和生物学作用尚未明确。为了获取有关AT2基因组DNA结构和功能的更多信息,我们克隆了小鼠AT2基因,并检测了其表达、转录和基因组结构。小鼠AT2 cDNA的氨基酸序列与大鼠AT2的序列一致性为98.5%。在小鼠胎儿中,AT2的mRNA在内脏和脑中高表达。出生后这种表达迅速下降。在10周龄小鼠中,通过Northern印迹分析可在脑中检测到AT2的mRNA。然而,逆转录-聚合酶链反应表明,在所检测的所有器官中均有AT2的mRNA表达,这表明AT2在其他器官中低水平表达。用BamHI、EcoRI和HindIII消化小鼠肝脏基因组DNA后的Southern印迹分析产生单一条带,表明AT2基因可能在小鼠基因组中以单一位点存在。AT2基因(EcoRI片段的4.5 kb)的核苷酸序列显示存在三个外显子。完整的编码序列包含在第三个外显子中。引物延伸实验表明小鼠AT2基因中存在两个转录起始位点。将小鼠AT2基因启动子区域(-1497至+56 bp)约1.5 kb的DNA片段与荧光素酶报告基因融合。(摘要截短至250字)

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