MOLECULAR CLONING OF BOMBESIN RECEPTOR

这个项目的目标是利用分子遗传技术来研究 哺乳动物蛙皮素受体的结构和功能。我们计划 通过首先获得鼠的cDNA克隆来实现该目的 在Swiss 3T3成纤维细胞上发现的蛙皮素受体相对较高， 号码将对cDNA进行测序，以确定预测的氨基酸 序列和受体的一般结构特征。 具体 将产生抗合成肽的抗血清， 受体蛋白的表征。然后可以使用该cDNA克隆 在位点特异性诱变研究中， 在突变受体表达后，可以检查干扰， 非洲爪蟾卵母细胞及DNA介导的小鼠成纤维细胞转染 这些细胞通常不表达受体。基因组克隆将是 分离以检查基因结构和启动子区域。的cDNA 编码区作为低严谨性探针筛选cDNA 从适当的来源，为其他不同的，但结构上的库 相关的铃蟾肽受体cDNA锥，包括一个独特的亚型，发现 在食道平滑肌和大脑嗅球上。

The goal of this project is to use molecular genetic techniques to study the structure and function of mammalian bombesin receptors. We plan to accomplish this objective by first obtaining cDNA clone for the murine bombesin receptor found on Swiss 3T3 fibroblasts in relatively high numbers. The cDNA will be sequenced to determine the predicted amino acid sequence and general structural features of the receptor. Specific antisera will be generated against synthetic peptides to allow further characterization of the receptor protein. This cDNA clone can then be used in site-specific mutagenesis studies where the effects of structural perturbations can be examined after expression of the mutant receptors in Xenopus oocytes and DNA-mediated transfection into murine fibroblasts which do not normally express the receptor. Genomic clones will be isolated to examine the gene structure and promoter region. The cDNA coding domain will be used as a low-stringency probe to screen cDNA libraries from appropriate sources for other distinct but structurally related bombesin receptor cDNA cones, including a distinct subtype found on esophageal smooth muscle and in the olfactory bulb of the brain.