Structural analysis of aldolase C gene and the identification of its expressed cell in brain

醛缩酶C基因的结构分析及其脑内表达细胞的鉴定

• 批准号: 62570116
• 负责人: MUKAI Tsunehiro
• 金额: $ 1.22万
• 依托单位: National Cardiovascular Center Research Institute (1988)佐賀医科大学 (1987)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62570116/
• 关键词: aldolase C; Purkinje cell; In Situ hybridization; in situ hybridization; 脳型アルドラーゼ; ID配列; In Situ Hybridization

Rat aldolase C gene was isolated. The gene comprises 9 exons interruped by 8 introns. The nucleotide sequence of the gene was determined, which includes 801 nucleotides(nt.) of 5'flanking, 3590 nt. of the gene region and 461 nt. of 3' flanking sequences. Two transcriptional initiation sites were also determined by the method of S1 mapping and primer extension. Aldolase C mRNA was exclusively expressed in brain and some in hepatoma. To identify the cell expressing aldolase C mRNA in brain we performed in situ hypridization of brain and spinal cord using riboprobe. We searched cerebral cortex, cerebellum, hippocampus, thalamus of the brain. Surprisingly silver grains accumulated in the Purkinje cell of the cerebellum and some in dorsal root ganglion and Nuc. motorius lateralis of the spinal cord. We also measured the copy number of the gene in the genome. From the measurement of the geme, titrated by Southern blotting, we conclude that a single copy of the rat aldolase C gene occurs per rat haploid genome.

分离大鼠醛缩酶C基因。该基因由9个外显子和8个内含子组成。测定了该基因的核苷酸序列，该基因包括801个核苷酸（nt.）5 '侧翼区3590 nt.的基因区域和461 nt. 3'侧翼序列。利用S1作图和引物延伸的方法确定了两个转录起始位点。醛缩酶C mRNA仅在脑组织中表达，在肝癌组织中也有部分表达。为了鉴定脑内表达醛缩酶C mRNA的细胞，我们用核糖核酸探针对脑和脊髓进行了原位杂交。我们搜索了大脑皮层，小脑，海马，丘脑。令人惊讶的是，银颗粒聚集在小脑浦肯野细胞和一些背根神经节和Nuc。脊髓的外侧运动肌我们还测量了基因组中基因的拷贝数。从测量的geme，滴定Southern印迹，我们得出结论，一个单一的副本，大鼠醛缩酶C基因发生每个大鼠单倍体基因组。

项目成果

K. Hori: "Isozymes. Structure and expression of human and rat isozyme gene:Multiple mRNA species of aldolase from a single gene" ALAN R. LISS. Inc., 23 (1987)

K. Hori：“同工酶。人类和大鼠同工酶基因的结构和表达：来自单个基因的醛缩酶的多种 mRNA 种类”ALAN R. LISS。

向井常博: "先天代謝異常の分子機構" 共立出版, 652-656 (1988)

Tsunehiro Mukai：“先天性代谢缺陷的分子机制”Kyoritsu Shuppan，652-656（1988）

T. Mukai.: "Structure and genomic organization of the brain-specific aldolase C gene"

T. Mukai.：“大脑特异性醛缩酶 C 基因的结构和基因组组织”

A. Kukita.: "The structure of brain-specific rat aldolase C mRNA and the evolution of aldolase isozyme genes" Eur. J. Biochem.171. 471-478 (1988)

A. Kukita.：“脑特异性大鼠醛缩酶 C mRNA 的结构和醛缩酶同工酶基因的进化”Eur。

H. Kishi: Pro. Natl. Acad. Sci. U. S. A.84. 8623-8627 (1987)

H.岸：赞成。