Chung S T
Bioprocess Research and Development, Upjohn Company, Kalamazoo, Michigan 49001.
J Bacteriol. 1987 Oct;169(10):4436-41. doi: 10.1128/jb.169.10.4436-4441.1987.
A 6.8-kilobase-pair (kbp) transposable element (Tn4556) was found in a neomycin-producing strain of Streptomyces fradiae. This element was first observed in two 30.3-kbp plasmids (pUC1123 and pUC1124) which arose when a thiostrepton resistance gene (1 kbp) was ligated with the BclI-2 fragment (22.5 kbp) that contains the origin of replication of phage SF1. The Tn4556 segment was deleted when these plasmids were transduced into another S. fradiae host with phage SF1. These deletion plasmids (pUC1210 and pUC1211) had copy numbers of less than 1 per chromosome and were unstable. In contrast, pUC1123 and pUC1124, with copy numbers of 12 to 15 per chromosome, respectively, were relatively stable. When pUC1210 and pUC1211 were reintroduced into S. fradiae by protoplast transformation, the Tn4556 element transposed again to the plasmids at numerous new locations in either of two orientations. A copy of Tn4556 was found in the S. fradiae chromosome by hybridization studies. It appears that Tn4556 originated from the chromosome, transposed into unstable pUC1210 and pUC1211, and made stable plasmids. A temperature-sensitive hybrid plasmid carrying a viomycin resistance derivative of Tn4556 (pMT660::Tn4556::vph) was constructed. When Streptomyces lividans UC8390 containing the hybrid plasmid was grown at 39 degrees C, Tn4556::vph (Tn4560) transposed to random positions in the host chromosome.
在弗氏链霉菌的新霉素生产菌株中发现了一个6.8千碱基对(kbp)的转座元件(Tn4556)。该元件最初在两个30.3 kbp的质粒(pUC1123和pUC1124)中观察到,这两个质粒是在硫链丝菌素抗性基因(1 kbp)与包含噬菌体SF1复制起点的BclI - 2片段(22.5 kbp)连接时产生的。当这些质粒用噬菌体SF1转导到另一个弗氏链霉菌宿主中时,Tn4556片段被删除。这些缺失质粒(pUC1210和pUC1211)每条染色体的拷贝数少于1个且不稳定。相比之下,pUC1123和pUC1124每条染色体的拷贝数分别为12至15个,相对稳定。当通过原生质体转化将pUC1210和pUC1121重新引入弗氏链霉菌时,Tn4556元件以两种方向中的任何一种再次转座到质粒的许多新位置。通过杂交研究在弗氏链霉菌染色体中发现了一个Tn4556拷贝。似乎Tn4556起源于染色体,转座到不稳定的pUC1210和pUC1121中,并形成了稳定的质粒。构建了一个携带Tn4556的紫霉素抗性衍生物的温度敏感型杂交质粒(pMT660::Tn4556::vph)。当含有该杂交质粒的淡紫灰链霉菌UC8390在39℃下生长时,Tn4556::vph(Tn4560)转座到宿主染色体的随机位置。