Shaw J H, Clewell D B
J Bacteriol. 1985 Nov;164(2):782-96. doi: 10.1128/jb.164.2.782-796.1985.
Streptococcus faecalis transposon Tn917 was cloned in Escherichia coli on plasmid vector pBR325. The erythromycin resistance determinant of Tn917 was not expressed in the E. coli background. The nucleotide sequence of Tn917 was determined and found to be 5,257 base pairs in length. Six open reading frames (ORFs) were identified and designated 1 through 6 (5' to 3'); all were on the same DNA strand. A region exhibiting strong homology with known promoters was identified upstream from ORF1. ORFs 1 to 3 were virtually identical to the previously sequenced erythromycin resistance determinant on Streptococcus sanguis plasmid pAM77. At the 3' point, where the homology between Tn917 and pAM77 ends, was a 20-base-pair region about 80% homologous with a component of the res site of Tn3. The amino acid sequence of ORF4 showed homology with other site-specific recombination enzymes, including approximately 30% homology with the resolvase of Tn3. Contained within Tn917 was a directly oriented 73-base-pair duplication of the left terminus. The Tn917 sequence revealed that antibiotic-enhanced transposition might be due to extension of transcription from the resistance-related genes (in ORFs 1 to 3) into transposition genes (in ORFs 4 to 6). Transcription analyses resulted in data consistent with this interpretation.
粪肠球菌转座子Tn917在大肠杆菌中克隆于质粒载体pBR325上。Tn917的红霉素抗性决定簇在大肠杆菌背景中不表达。测定了Tn917的核苷酸序列,发现其长度为5257个碱基对。鉴定出6个开放阅读框(ORF),分别命名为1至6(5'至3');所有这些都位于同一条DNA链上。在ORF1上游鉴定出一个与已知启动子具有高度同源性的区域。ORF1至3与先前测序的血链球菌质粒pAM77上的红霉素抗性决定簇几乎相同。在Tn917和pAM77之间同源性结束的3'端,是一个与Tn3的res位点的一个组分约80%同源的20个碱基对的区域。ORF4的氨基酸序列与其他位点特异性重组酶显示出同源性,包括与Tn3的解离酶约30%的同源性。Tn917内包含左末端的一个直接定向的73个碱基对的重复序列。Tn917序列表明,抗生素增强的转座可能是由于转录从抗性相关基因(在ORF1至3中)延伸到转座基因(在ORF4至6中)。转录分析得到的数据与这一解释一致。