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一个孤儿阿片受体基因家族成员及其剪接变体的cDNA克隆

cDNA cloning of an orphan opiate receptor gene family member and its splice variant.

作者信息

Wang J B, Johnson P S, Imai Y, Persico A M, Ozenberger B A, Eppler C M, Uhl G R

机构信息

Molecular Neurobiology Branch, National Institute on Drug Abuse, NIH, Baltimore, MD.

出版信息

FEBS Lett. 1994 Jul 4;348(1):75-9. doi: 10.1016/0014-5793(94)00557-5.

Abstract

Radioligand binding and cDNA homology studies have suggested the existence of opiate receptors distinct from the recently-cloned mu, delta and kappa receptors. XOR1S, a rat brain cDNA whose predicted translation product displays 67-72% homology with those encoded by mu 1, delta 1 and kappa 1 opiate receptor cDNAs, was constructed from two partial cDNAs identified through cDNA homology approaches. A longer XOR1L variant of this cDNA was also identified by polymerase chain reaction studies using genomic DNA and cDNA from brain and peripheral tissues. XOR1 mRNA is most highly expressed in hypothalamus. COS cell expression of both clones confers neither robust binding of opiate ligands nor reproducible opiate inhibition of forskolin-stimulated adenylate cyclase. These studies identify an orphan clone that helps to define features of the opiate receptor gene family, including apparent differential splicing and expression in peripheral tissues.

摘要

放射性配体结合和cDNA同源性研究表明,存在与最近克隆的μ、δ和κ受体不同的阿片受体。XOR1S是一种大鼠脑cDNA,其预测的翻译产物与由μ1、δ1和κ1阿片受体cDNA编码的产物具有67 - 72%的同源性,它由通过cDNA同源性方法鉴定的两个部分cDNA构建而成。通过使用来自脑和外周组织的基因组DNA和cDNA进行聚合酶链反应研究,还鉴定出了该cDNA的一个更长的XOR1L变体。XOR1 mRNA在下丘脑中表达最高。这两个克隆在COS细胞中的表达既没有赋予阿片配体强大的结合能力,也没有对福斯可林刺激的腺苷酸环化酶产生可重复的阿片抑制作用。这些研究鉴定出了一个孤儿克隆,它有助于定义阿片受体基因家族的特征,包括明显的差异剪接和在外周组织中的表达。

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