Heuer C, Hickman R K, Curiale M S, Hillen W, Levy S B
J Bacteriol. 1987 Mar;169(3):990-4. doi: 10.1128/jb.169.3.990-994.1987.
The Tn10-like constitutively expressed tetracycline resistance determinant from a Haemophilus parainfluenzae strain was cloned in Escherichia coli. Toxicity resulting from expression on multicopy plasmids necessitated its being cloned on a low-copy plasmid vector or in cells containing the Tn10-encoded repressor. Constitutive expression of tetracycline resistance was found to result from the synthesis of a truncated inactive repressor molecule. Instead of the 23-kilodalton repressor found in other Tn10-containing strains, this determinant encoded a 14.5-kilodalton molecule. The DNA sequence of the 700-base-pair region spanning the repressor gene and promoter-operator regions of the Haemophilus determinant was identical to that of the same region of Tn10, except for the absence of a single T X A base pair in the repressor gene. This deletion leads to premature termination of the protein. Antisera to the repressor suggested that the repressor was also absent in a second independently isolated H. parainfluenzae strain bearing a Tn10-like constitutive tetracycline resistance determinant.
从副流感嗜血杆菌菌株中分离出的类Tn10组成型表达的四环素抗性决定簇在大肠杆菌中进行了克隆。多拷贝质粒上表达所产生的毒性使得其必须克隆到低拷贝质粒载体上,或克隆到含有Tn10编码阻遏物的细胞中。发现四环素抗性的组成型表达是由截短的无活性阻遏物分子的合成导致的。该决定簇编码的不是在其他含Tn10菌株中发现的23千道尔顿的阻遏物,而是一个14.5千道尔顿的分子。跨越嗜血杆菌决定簇阻遏基因以及启动子-操纵子区域的700碱基对区域的DNA序列,与Tn10相同区域的序列一致,只是阻遏基因中缺少一个T X A碱基对。这种缺失导致蛋白质提前终止。针对阻遏物的抗血清表明,在另一株独立分离的携带类Tn10组成型四环素抗性决定簇的副流感嗜血杆菌菌株中也不存在该阻遏物。