Secretion-dependent proteolysis of recombinant proteins is associated with inhibition of cell growth in Escherichia coli
Secretion-dependent proteolysis of recombinant proteins is associated with inhibition of cell growth in Escherichia coli
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DOI：
10.1023/a:1018363203858
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发表时间：
1997-04-01
影响因子：
2.7
通讯作者：
Villaverde, A
中科院分区：
文献类型：
--
作者：
Viaplana, E;Rebordosa, X;Villaverde, A
在IPTG诱导的T7启动子控制下，在大肠杆菌中表达兔出血症病毒VP60衣壳蛋白的抗原性C末端。设计了两个不同但密切相关的结构，在病毒片段的N端携带周质分泌信号或T7检测标签。细胞质蛋白的产量很高，而周质蛋白在Western印迹中很难检测到，因为它在合成后立即降解。产生周质形式而不是细胞质形式的重组培养物在诱导基因表达后显示细胞生长急剧停止，这表明与重组蛋白本身或其蛋白分解过程有关的毒性。对这种毒性效应的分子机制进行了讨论。
The antigenic C-terminus of VP60 capsid protein from rabbit haemorrhagic disease virus was produced in E. coli under the control of an IPTG-inducible T7 promoter. Two different but closely related constructs were designed, carrying either a periplasmic secretional signal or a T7 detection tag at the N-terminus of the viral segment. The cytoplasmic protein is produced in high yields whereas the periplasmic version is hardly detected in Western blot, due to its immediate degradation after synthesis. Recombinant cultures producing the periplasmic, but not the cytoplasmic form show a dramatic arrest of cell growth after induction of gene expression, indicative of toxicity associated to the recombinant protein itself or to its proteolytic processing. Molecular mechanisms for such toxic effects are discussed.