Kim E H, Aoki T
Department of Biological Resources, Faculty of Agriculture, Miyazaki University, Japan.
Microbiol Immunol. 1994;38(1):31-8. doi: 10.1111/j.1348-0421.1994.tb01741.x.
Tetracycline (pp-tet), and kanamycin (pp-kan) resistance genes were cloned from a transferable R plasmid of fish pathogen Pasteurella piscicida, and complete nucleotide sequences were determined. The pp-tet was a class D Tet determinant constructed with the tetA resistance gene of 1,182 bp encoding a protein with a deduced molecular mass of 41 kDa and the tetR repressor gene of 654 bp encoding a product of 24 kDa. The pp-tet was highly homologous to the tet(D) of plasmid RA1 isolated from Aeromonas hydrophila with two nucleotide differences in the tetR, and of plasmid pIP173 from Salmonella ordonez with two nucleotide differences in the tetA. The pp-kan contained 813 bp encoding a 31 kDa protein of 271 amino acids, and was classified into type aph-Ic. It was identical to the aphA7 in the IAB operon of pBWH77, in which was originally found an isolate of Klebsiella pneumoniae, in its nucleotide sequences and hybrid promoter construction. The genes were connected by an insertion sequence IS26 of 820 bp, and were flanked by repeated copies in direct orientation at the 3' flanking region of the pp-tetA and in inverted orientation at the 3' flanking region of the pp-kan. The genetic elements are organized like a complex transposon by close linkage of the IS26 and the pp-tet and -kan.
从鱼类病原菌杀鲑气单胞菌的可转移R质粒中克隆了四环素(pp - tet)和卡那霉素(pp - kan)抗性基因,并测定了其完整的核苷酸序列。pp - tet是一种D类四环素决定簇,由1182 bp的tetA抗性基因构建而成,该基因编码一种推导分子量为41 kDa的蛋白质,还有654 bp的tetR阻遏基因,编码一种24 kDa的产物。pp - tet与从嗜水气单胞菌分离的质粒RA1的tet(D)高度同源,tetR中有两个核苷酸差异;与来自肠炎沙门氏菌的质粒pIP173的tet(D)也高度同源,tetA中有两个核苷酸差异。pp - kan包含813 bp,编码一个由271个氨基酸组成的31 kDa蛋白质,被归类为aph - Ic型。它在核苷酸序列和杂交启动子结构上与pBWH77的IAB操纵子中的aphA7相同,pBWH77最初是从肺炎克雷伯菌的一个分离株中发现的。这些基因由一个820 bp的插入序列IS26连接,在pp - tetA的3'侧翼区域以正向重复拷贝为侧翼,在pp - kan的3'侧翼区域以反向重复拷贝为侧翼。这些遗传元件通过IS26与pp - tet和 - kan的紧密连锁,像一个复合转座子一样组织起来。