NOVEL HIGH-LEVEL EXPRESSION CLONING VEHICLES - 10(4)-FOLD AMPLIFICATION OF ESCHERICHIA-COLI MINOR PROTEIN
NOVEL HIGH-LEVEL EXPRESSION CLONING VEHICLES - 10(4)-FOLD AMPLIFICATION OF ESCHERICHIA-COLI MINOR PROTEIN
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DOI：
10.1038/nbt0184-81
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发表时间：
1984-01-01
期刊：
影响因子：
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通讯作者：
NOUYE, M
中科院分区：
文献类型：
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作者：
MASUI, Y;MIZUNO, T;NOUYE, M
通用的、高水平的表达克隆载体pINIII已经被构建。外源DNA片段可以插入起始密码子之后的唯一EcoRI、HindIII或BamHI位点上的三个阅读框中的任何一个。克隆的外源基因同时受启动子和启动子操纵子的控制。该基因的表达受同一载体产生的抑制物的调节。利用pINIII载体EnvZ蛋白，大肠杆菌的一个非常微小的调节蛋白被扩增了104倍以上，占总细胞蛋白的30%。
Versatile, high-level expression cloning vehicles, pINIII, have been constructed. A foreign DNA fragment can be inserted in any one of the three reading frames at the unique EcoRI, HindIII, or BamHI site immediately after the initiation codon. The cloned foreign gene is under the control of both thelpppromoter and thelacpromoter-operator. The expression of the gene is regulated by thelacrepressor produced by the same vehicles. Using a pINIII vehicle EnvZ protein, a very minor regulatory protein ofEscherichia coliwas amplified more than 104-fold, becoming 30% of total cellular protein.