Nagai M, Hiramatsu R, Kanéda T, Hayasuke N, Arimura H, Nishida M, Suyama T
Gene. 1985;36(1-2):183-8. doi: 10.1016/0378-1119(85)90084-8.
A cDNA library was constructed in pBR322 from 18S to 20S mRNA that was extracted from human kidney cells, fractionated on oligo(dT)-cellulose column and sucrose-density gradient, and confirmed for urokinase production in Xenopus laevis oocytes. The Escherichia coli RR1 transformants were hybridized to synthetic oligonucleotide probe prepared according to the known amino acid sequence, Glu 73 to Glu 77 of human urinary urokinase chain B. The entire cloned cDNA covers a 2250-bp region, wherein the 1293-bp sequence codes for preprourokinase consisting of 431 amino acids, with the first 20 residues being a signal peptide. The 5'-untranslated region is at least 80 bp long and the 3'-untranslated region is longer than 850 bp.
从人肾细胞中提取18S至20S的mRNA，在寡聚（dT）-纤维素柱和蔗糖密度梯度上分级分离，然后构建于pBR322中的cDNA文库，并在非洲爪蟾卵母细胞中证实其产生尿激酶。将大肠杆菌RR1转化体与根据人尿激酶B链已知氨基酸序列Glu 73至Glu 77制备的合成寡核苷酸探针杂交。整个克隆的cDNA覆盖2250bp区域，其中1293bp序列编码由431个氨基酸组成的前尿激酶原，前20个残基为信号肽。5'-非翻译区长至少80bp，3'-非翻译区长超过850bp。
