Production of human renin in microorganism and its application to medicine.

微生物生产人肾素及其在医药上的应用。

• 批准号: 58440078
• 负责人: MURAKAMI Kazuo
• 金额: $ 19.65万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1983
• 资助国家: 日本
• 起止时间: 1983 至 1985
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-58440078/
• 关键词: Renin; Prorenin; cDNA; 大腸菌; 発現

1) The primary structure of human renin precursor was deduced from its complementary sequence. The predicted sequence consists of 406 amino acids with a pre- and a prosegment carrying 20 and 46 amino acids, respectively.2) A three-dimensional model of human renin has been constructed based on the assumption that the overall folding of the aspartyl proteases is very similar. As a reference, we used penicillopepsin.3) The human renin gene was isolated from a Charon 4A human genomic library and characterized. The gene spans about 11.7 kilobases and consists of 10 exons and 9 introns that map at points that could be variable surface loops of the enzyme.4) Human prorenin was produded under the transcriptional control of the E. coli trp promoter-operator in the expression vector. The transformed E. coli cell expressed high levels (30% of total cell protein) of prorenin as part of a hybrid protein.

1)根据人肾素前体的互补序列推测其一级结构。预测的序列由406个氨基酸组成，前段和前段分别由20个和46个氨基酸组成。2)基于天冬氨酸蛋白酶的整体折叠非常相似的假设，构建了人肾素的三维模型。3)从Charon 4A人类基因组文库中分离并鉴定了人肾素基因。该基因全长约11.7kb，由10个外显子和9个内含子组成，这些外显子和9个内含子映射在可能是酶的可变表面环的点上。4)人Prorenin基因是在表达载体中的E.coliTrp启动子-操纵子的转录控制下产生的。转化的大肠杆菌细胞表达了高水平(占总细胞蛋白的30%)的原蛋白，作为杂交蛋白的一部分。

项目成果

Hypertension. Vol. 7. (1985)

高血压。

Proc. Natl. Acad. Sci. USA. Vol. 81. (1984)

过程。

Hypertension. 7-5. (1985)

高血压。

The Journal of Biological Chemistry. Vol. 260. (1985)

生物化学杂志。

Proc. Natl. Acad. Sci. USA. Vol. 80. (1983)

过程。