THE MOLECULAR BIOLOGY OF THE MAMMALIAN GRP GENE FAMILY

哺乳动物前胃泌素原的结构、功能和表达 释放肽（GRP）基因的研究。 实验 a）表征生物体的结构和调节， 大鼠前原GRP基因，B）过表达人GRP原激素 分析蛋白质翻译后加工和肽 激素功能，c）获得其它铃蟾肽家族cDNA克隆 激素原基因 A.大鼠前原GRP基因。 结构和整个核苷酸 使用基因组测序法测定大鼠前原GRP基因的序列。 克隆和从大鼠脑cDNA文库分离的cDNA克隆。 在 与人类基因相比， 在表达该基因的大鼠组织中观察到。 编码区 因为GRP及其相关肽在进化上是保守的， 与两个领域的生物学功能一致。 大鼠GRP 基因由脑中的组织特异性启动子转录， 创造了另一种调节基因表达的途径。 的 脑特异性启动子仅在大鼠的选定核中使用 通过原位杂交在细胞水平上分析时，大脑。 B。人前原GRP基因成功地在一个细胞中过表达。 新型杆状病毒蛋白表达系统。 利用该模型 系统，可以产生足够的蛋白质以允许分析 翻译后加工事件需要生成 生物活性肽，并纯化这些肽， 建立功能。 C.我们设计并合成了共有寡核苷酸， 特异于受体结合生物活性的探针 蛙皮素家族肽的羧基七肽。 这 寡核苷酸探针目前用于获得和 表征在神经细胞中表达的其他铃蟾肽家族基因 组织.

The structure, function, and expression of mammalian prepro gastrin releasing peptide (GRP) gene was studied. Experiments were performed to a) characterize the structure and regulation of the rat prepro GRP gene, b) overexpress the human GRP prohormone protein to analyze post-translational processing and peptide hormone function, c) obtain cDNA clones for other bombesin family prohormone genes. A. Rat prepro GRP gene. The structure and entire nucleotide sequence of the rat prepro GRP gene was determined, using genomic clones and cDNA clones isolated from a rat brain cDNA library. In contrast to the human gene, there is no alternative RNA processing observed in rat tissues expressing this gene. The coding regions for GRP and its associated peptide are evolutionarily conserved, consistent with a biologic function for both domains. The rat GRP gene is transcribed from a tissue specific promoter in the brain, creating an alternative route for regulating gene expression. The brain specific promoter is used only in selected nuclei in the rat brain when analyzed at the cellular level by in situ hybridization. B. The human prepro GRP gene was successfully overexpressed in a novel baculovirus protein expression system. Using this model system, sufficient protein can be produced to allow analysis of post-translational processing events needed to generate biologically active peptides, and purify those peptides to establish function. C. We have designed and synthesized consensus oligonucleotide probes specific for the receptor-binding biologically active carboxyl heptapeptide of bombesin family peptides. This oligonucleotide probe is currently being used to obtain and characterize other bombesin family genes expressed in neural tissue.