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Molecular cloning and chromosomal localization of the genes encoding P23k neuron-specific calcium-binding protein family.

编码 P23k 神经元特异性钙结合蛋白家族的基因的分子克隆和染色体定位。

基本信息

批准号：
08670733
负责人：
TAKAMATSU Ken
金额：
$ 1.22万
依托单位：
Toho University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670733/
关键词：
Hippocalcin Calcium-binding protein Brain Hippocampus Gene Chromosomallocalization 染色体

项目摘要

P23k family is a novel EF-hand calcium-binding protein subfamily, considered to be a calcium sensor in neuron of the central nervous system. We initiated a search for the brain member of P23k family in human and mouse. We report here on two novel human cDNAs and mouse hippocalcin gene.Two novel human cDNAs, HLP3 and HLP4, were obtained from a human brain cDNA library. Both cDNA encode a homologous protein with a molecular weight of 23kDa, containing four potential EF-hand motifs. Northern blot analysis revealed that both mRNAs were expressed only in the brain. The HLP3 mRNA was distributed to most of the brain regions, high amounts of the HLP3mRNA were detected in the cerebral cortex and cerebellum. The HLP4mRNA was also found in many regions of the brain, relatively high amounts of the HLP4mRNA were detected in the cerebral cortex and cerebellum. By discordancy analysis using DNAs from human-rodent somatic cell hybrids, the HLP3 and HLP4 genes were mapped to chromosome 2 and 17, respectively.The mouse hippocalcin gene was obtained from a mouse leukocyte genomic library. The mouse hippocalcin gene contains three exons and two introns, which spans approximately 8 kb. Deduced amino acid sequence of mouse hippocalcin is 100% identical to those of the rat and human and structures of hippocalcin genes are highly conserved across species. The transcription termination site was assigned at 860bp from TAG.The multiple transcription initiation sites were assigned at the positions around-180bp. Neither a canonical 'TATA' box, and 'CAAT' box was found in the 5'-flanking region. By Southern blot analysis, the positive bands exactlycoincided with those expected from the cloned gene. Fluorescence in situ hybridization revealed the mouse hippocalcin gene mapped to chromosome 4, region D2-D3.

P23k家族是一个新的EF-手钙结合蛋白亚家族，被认为是中枢神经系统神经元中的钙感受器。我们开始在人和小鼠中寻找P23k家族的脑成员。我们报道了两个新的人和小鼠海马素基因，其中两个新的人基因HLP3和HLP4是从人脑cDNA文库中获得的。这两个基因都编码一个分子量为23 kDa的同源蛋白，包含四个潜在的EF-Hand基序。Northern印迹分析表明，这两种mRNAs仅在脑内表达。HLP3mRNA分布于大部分脑区，在大脑皮层和小脑中表达较高。HLP4mRNA也存在于大脑的许多区域，在大脑皮层和小脑中检测到相对较高的HLP4mRNA。通过对人-鼠体细胞杂交DNA的不一致性分析，将HLP3和HLP4基因分别定位在第2号和第17号染色体上。小鼠海马素基因由3个外显子和2个内含子组成，全长约8kb。小鼠海马素的推导氨基酸序列与大鼠和人的氨基酸序列100%相同，并且海马素基因的结构在物种之间高度保守。转录终止位点位于TAG的860bp处，多个转录起始点位于-180bp左右。在5‘侧翼区没有发现典型的’TATA‘盒和’CAAT‘盒。经Southern杂交分析，阳性条带与所克隆基因的预期条带完全一致。荧光原位杂交显示小鼠海马素基因定位于4号染色体D2-D3区域。