Kashiwagi K, Suzuki T, Suzuki F, Furuchi T, Kobayashi H, Igarashi K
Faculty of Pharmaceutical Sciences, Chiba University, Japan.
J Biol Chem. 1991 Nov 5;266(31):20922-7.
The nucleotide sequence of one of the putrescine transport operons (pPT71), located at 16 min of the Escherichia coli chromosome, was determined. It contained the genes for an induced ornithine decarboxylase and a putrescine transport protein. The gene for the ornithine decarboxylase contained a 2,196-nucleotide open reading frame encoding a 732-amino acid protein whose calculated Mr was 82,414, and the predicted amino acid sequence from the open reading frame had 65% homology with that of a constitutive ornithine decarboxylase encoded by the gene at 64 min. The ornithine decarboxylase activity was observed in the cells carrying pPT71 cultured at pH 5.2, but not in the cells cultured at pH 7.0. The gene for the putrescine transport protein contained a 1,317-nucleotide open reading frame encoding a 439-amino acid protein whose calculated Mr was 46,494. The hydropathy profile of the putrescine transport protein revealed that it consisted of 12 putative transmembrane spanning segments linked by hydrophilic segments of variable length. The transport protein was in fact found in the membrane fraction. When the gene for the putrescine transport protein was linked to the tet promoter of the vector instead of its own promoter, the putrescine transport activity increased greatly. The results suggest that the gene expression of the operon is repressed strongly under standard conditions.
确定了位于大肠杆菌染色体16分钟处的一个腐胺转运操纵子(pPT71)的核苷酸序列。它包含诱导型鸟氨酸脱羧酶和腐胺转运蛋白的基因。鸟氨酸脱羧酶基因包含一个2196个核苷酸的开放阅读框,编码一个732个氨基酸的蛋白质,其计算的Mr为82414,并且该开放阅读框预测的氨基酸序列与位于64分钟处的基因编码的组成型鸟氨酸脱羧酶的氨基酸序列具有65%的同源性。在pH 5.2培养的携带pPT71的细胞中观察到鸟氨酸脱羧酶活性,但在pH 7.0培养的细胞中未观察到。腐胺转运蛋白基因包含一个1317个核苷酸的开放阅读框,编码一个439个氨基酸的蛋白质,其计算的Mr为46494。腐胺转运蛋白的亲水性图谱显示它由12个推定的跨膜区段组成,这些区段由可变长度的亲水性区段连接。实际上在膜组分中发现了转运蛋白。当腐胺转运蛋白基因与载体的tet启动子相连而不是其自身的启动子时,腐胺转运活性大大增加。结果表明该操纵子的基因表达在标准条件下受到强烈抑制。