Finn C W, Silver R P, Habig W H, Hardegree M C, Zon G, Garon C F
Science. 1984 May 25;224(4651):881-4. doi: 10.1126/science.6326263.
A pool of synthetic oligonucleotides was prepared based on the amino terminal amino acid sequence of tetanus toxin. This probe hybridized to plasmid DNA isolated from three toxigenic strains of Clostridium tetani but not to plasmid DNA from a nontoxigenic strain. These results show that the structural gene for the toxin is on the plasmid. The pCL1 plasmid from one of the toxigenic strains spontaneously deleted 22 kilobase pairs of DNA to form pCL2. Strains harboring this deleted plasmid are nontoxigenic. However, the probe mixture hybridized to pCL2, indicating that the DNA encoding the amino terminus of the toxin had not been deleted. Restriction endonuclease cleavage maps of pCL1 and pCL2 were constructed and indicate the approximate location and orientation of the structural gene for tetanus toxin.
基于破伤风毒素的氨基末端氨基酸序列制备了一组合成寡核苷酸。该探针与从三株产毒破伤风梭菌分离的质粒DNA杂交,但不与非产毒菌株的质粒DNA杂交。这些结果表明毒素的结构基因位于质粒上。来自其中一株产毒菌株的pCL1质粒自发缺失了22千碱基对的DNA,形成了pCL2。携带这种缺失质粒的菌株不产毒。然而,探针混合物与pCL2杂交,表明编码毒素氨基末端的DNA未被缺失。构建了pCL1和pCL2的限制性内切酶切割图谱,显示了破伤风毒素结构基因的大致位置和方向。