MOLECULAR BIOLOGICAL STUDIES OF CALRETININ

为了探讨Calretinin在大脑中的作用及其在脑内的作用 在分子水平上的调控，我们克隆并分析了大鼠 Calretinin基因的编码区。从一株小鼠中分离到一个免疫反应克隆 大鼠脑表达文库lambda-GT 11.1.45kb的插入片段 被亚克隆到pGEM-4Z转录载体的Eco RI位点上 以作进一步分析。它的身份是通过与 人钙调蛋白。大鼠的cDNA5‘端序列为54bp5’未翻译 一个816bp的开放阅读框(ORF；)，包括开始和停止 密码子，以及一个579bp3‘非翻译区。ORF有271个密码子 编码一个31.4 kDa的假定蛋白质。多聚腺苷基化信号和 3‘端附近有13个腺苷残基。 进化保守的钙结合结构域及其连接 大鼠和雏鸡原代之间的区域和有限的变化 结构使我们相信Calretinin与其他高度相互作用的 脑细胞的保守成分。这个claretinin cdna克隆 为分析中枢特定神经元提供了一种新的探索。 神经系统。该cDNA探针将允许更详细地分析 Calretinin在大脑中的表达，将有助于筛查 完整染色体基因的基因组文库。

In order to explore the role of calretinin in the brain and its regulation at the molecular level, we have cloned and analyzed the rat cDNA coding for calretinin. An immunoreactive clone was isolated from a rat brain cDNA expression library in lambda-gt 11. The 1.45 kb insert was subcloned into the Eco RI site of the pGEM-4Z transcription vector for further analysis. Its identity was confirmed by comparison with human calretinin. The rat cDNA sequence comprised a 54 bp 5'untranslated region, an 816 bp open reading frame (ORF;) including start and stop codons, and a 579 bp 3' untranslated region. The ORF has 271 codons coding for a putative protein of 31.4 kDa. A polyadenylation signal and 13 adenylate residues were found near the 3' end. The evolutionarily conserved calcium binding domains and connecting regions and the limited changes observed between rat and chick primary structure lead us to believe that calretinin interacts with other highly conserved constituents of brain cells. This claretinin cDNA clone provides a new probe for the analysis of specific neurons in the central nervous system. The cDNA probe will allow a more detailed analysis of calretinin expression in the brain and will be useful for screening genomic libraries for the complete chromosomal gene.