ARTIFICIAL REGULATION OF GENE-EXPRESSION IN ESCHERICHIA-COLI BY RNASE-P
ARTIFICIAL REGULATION OF GENE-EXPRESSION IN ESCHERICHIA-COLI BY RNASE-P
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DOI：
10.1073/pnas.92.24.11115
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发表时间：
1995-11-21
影响因子：
11.1
通讯作者：
ALTMAN, S
中科院分区：
文献类型：
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作者：
GUERRIERTAKADA, C;LI, Y;ALTMAN, S
构建了编码各种外部指导序列（EGS）的质粒并插入大肠杆菌中。在携带适当质粒的菌株中，完全诱导的β-半乳糖苷酶和碱性磷酸酶活性的表达降低了50%以上，而在具有非特异性EgSs的菌株中没有观察到这种活性的降低。在限制性温度下，对RNase P具有温度敏感性的菌株中基因表达的抑制几乎被消除（EC 3.1.26.5）。北方印迹分析表明EGSRNA在体内的稳态拷贝数为几百个/细胞。一个含有M1 RNA基因的质粒共价连接到一个特定的EGS，降低了由一个单独的质粒编码的抑制tRNA的表达水平。类似的方法也可用于调控大肠杆菌中的基因表达。大肠杆菌，并模拟冷敏感突变体的性质。
Plasmids encoding various external guide sequences (EGSs) were constructed and inserted into Escherichia coli. In strains harboring the appropriate plasmids, the expression of fully induced beta-galactosidase and alkaline phosphatase activity was reduced by more than 50%, while no reduction in such activity was observed in strains with non-specific EgSs. The inhibition of gene expression was virtually abolished at restrictive temperatures in Strains that were temperature-sensitive for RNase P (EC 3.1.26.5). Northern blot analysis shelved that the steady-state copy number of EGS RNAs was several hundred per cell in vivo. A plasmid that contained a gene for M1 RNA covalently linked to a specific EGS reduced the level of expression of a suppressor tRNA that was encoded by a separate plasmid. Similar methods can be used to regulate gene expression in E. coli and to mimic the properties of cold-sensitive mutants.