Conditional expression of the vesicular stomatitis virus glycoprotein gene in Escherichia coli.
Conditional expression of the vesicular stomatitis virus glycoprotein gene in Escherichia coli.
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水泡性口炎病毒糖蛋白基因在大肠杆菌中的条件表达。
DOI：
10.1073/pnas.78.11.6670
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发表时间：
1981
影响因子：
11.1
通讯作者：
Shafferman,A
中科院分区：
文献类型：
--
作者：
Rose,JK;Shafferman,A
构建了在色氨酸操纵子调控区控制下指导水泡性口炎病毒糖蛋白（G蛋白）基因表达的细菌质粒。指导G蛋白样蛋白合成的质粒（含有由trpE基因编码的7个氨基酸的NH 2-末端片段，该片段与仅缺少其NH 2-末端甲硫氨酸的完整G蛋白序列融合）可以转化为trpR+（阻遏）细胞，但不能转化为trpR-（去阻遏）细胞。这一结果初步表明，由该质粒编码的G蛋白样蛋白的去阻遏合成在大肠杆菌中是致命的。缺失编码G蛋白COOH末端附近的大疏水片段的序列并不能克服这种致死性。去阻遏合成的致死性通过在疏水NH 2-末端结构域（信号肽）中缺失编码10个氨基酸的G蛋白基因区域来克服。胰蛋白酶肽图分析表明，该质粒编码的G蛋白样蛋白和一些截短蛋白含有G蛋白的蛋白序列。水泡性口炎病毒的抗血清沉淀的G蛋白样蛋白，表明它与真正的G蛋白共有抗原决定簇。
Bacterial plasmids that directed expression of the vesicular stomatitis virus glycoprotein (G-protein) gene under control of the tryptophan operon regulatory region were constructed. A plasmid directing the synthesis of a G-protein-like protein (containing the NH2-terminal segment of seven amino acids encoded by the trpE gene fused to the complete G-protein sequence lacking only its NH2-terminal methionine) could be transformed into trpR+ (repressed) but not into trpR- (derepressed) cells. This result suggested initially that derepressed synthesis of the G-protein-like protein encoded by this plasmid was lethal in Escherichia coli. Deletion of the sequence encoding the large hydrophobic segment near the COOH terminus of G-protein did not overcome this lethality. Lethality of derepressed synthesis was overcome by deletion of the G-protein gene region encoding 10 amino acids in the hydrophobic NH2-terminal domain (signal peptide). Tryptic peptide mapping demonstrated that the G-protein-like protein and some truncated proteins encoded by the plasmid contained G-protein protein sequences. Antisera to vesicular stomatitis virus precipitated the G-protein-like protein, showing that it shares antigenic determinants with the authentic G-protein protein.