Cloning of Neuron Specific Protein, MxR

神经元特异性蛋白 MxR 的克隆

• 批准号: 09670680
• 负责人: TSUBOI Yoshio
• 金额: $ 1.98万
• 依托单位: Fukuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670680/
• 关键词: Endoplasmic reticulum; Neuron; Neuroendocrine specific protein; Reticulon; cDNA cloning; FISH; Mx-1; マウス; 核; 軸原; シナクロス; 軸策; シナプシス

We cloned a cDNA encoding mouse neuroendocrine specific protein/reticulon (RTN) 3 by screening mice brain cDNA library. The cDNA of mRTN3/mMxR1 was 1745nts and contains 711nts or open reading frame (ORF). It has 832nls of long 3' noncoding region (NCR), and poly A signal (AATAAA) is located at l694. The sequence analysis indicated that mRTN3/mMxR1 has strong homology with human RTN in its C-terminus. Northern blot analysis indicated that mRTN3/mMxR1 was expressed in brain. Its expression was not controlled embryonic-developmentally. We expressed the ORF as a fusion protein with green fluorescent protein in COS 7 or HeLa cells. The expressed fusion protein was localized in the endoplasmic reticulum. When the unique N-terminal sequence was removed, the truncated fusion protein remained in the ER though the intensity of the protein was reduced. We are now trying to determine the topology of mRTN3/mMxR1 to the ER membrane. We are also trying to make a specific antibodies by immunizing rabbits with synthetic peptides.

我们通过筛选小鼠脑cDNA文库，克隆了编码小鼠神经内分泌特异性蛋白/网状蛋白（RTN）3的cDNA。 mRTN3/mMxR1的cDNA为1745nts，包含711nts或开放阅读框(ORF)。它有832nls的长3'非编码区(NCR)，多聚腺苷酸信号(AATAAA)位于l694。序列分析表明mRTN3/mMxR1的C端与人RTN具有很强的同源性。 Northern印迹分析表明mRTN3/mMxR1在脑中表达。其表达不受胚胎发育控制。我们在 COS 7 或 HeLa 细胞中将 ORF 表达为与绿色荧光蛋白的融合蛋白。表达的融合蛋白定位于内质网。当独特的 N 端序列被去除时，截短的融合蛋白仍保留在 ER 中，尽管蛋白质的强度降低了。我们现在正在尝试确定 mRTN3/mMxR1 到 ER 膜的拓扑结构。我们还尝试用合成肽对兔子进行免疫来制备特异性抗体。

项目成果

Kawaguchi et al.: "Rare tevelency of catching cold in Parkinson's disease"Parkinsonism and Related disordes. 4. 207-209 (1998)

Kawaguchi 等人：“帕金森病中罕见的感冒”帕金森病及相关疾病。