Jiang S C, Paul J H
Marine Science Department, University of South Florida, St. Petersburg, Florida 33701, USA.
Appl Environ Microbiol. 1998 Aug;64(8):2780-7. doi: 10.1128/AEM.64.8.2780-2787.1998.
To determine the potential for bacteriophage-mediated gene transfer in the marine environment, we established transduction systems by using marine phage host isolates. Plasmid pQSR50, which contains transposon Tn5 and encodes kanamycin and streptomycin resistance, was used in plasmid transduction assays. Both marine bacterial isolates and concentrated natural bacterial communities were used as recipients in transduction studies. Transductants were detected by a gene probe complementary to the neomycin phosphotransferase (nptII) gene in Tn5. The transduction frequencies ranged from 1.33 x 10(-7) to 5.13 x 10(-9) transductants/PFU in studies performed with the bacterial isolates. With the mixed bacterial communities, putative transductants were detected in two of the six experiments performed. These putative transductants were confirmed and separated from indigenous antibiotic-resistant bacteria by colony hybridization probed with the nptII probe and by PCR amplification performed with two sets of primers specific for pQSR50. The frequencies of plasmid transduction in the mixed bacterial communities ranged from 1.58 x 10(-8) to 3.7 x 10(-8) transductants/PFU. Estimates of the transduction rate obtained by using a numerical model suggested that up to 1.3 x 10(14) transduction events per year could occur in the Tampa Bay Estuary. The results of this study suggest that transduction could be an important mechanism for horizontal gene transfer in the marine environment.
为了确定噬菌体介导的基因转移在海洋环境中的可能性,我们利用海洋噬菌体宿主分离株建立了转导系统。含有转座子Tn5并编码卡那霉素和链霉素抗性的质粒pQSR50用于质粒转导试验。海洋细菌分离株和浓缩的天然细菌群落都被用作转导研究的受体。通过与Tn5中新霉素磷酸转移酶(nptII)基因互补的基因探针检测转导子。在用细菌分离株进行的研究中,转导频率范围为1.33×10^(-7)至5.13×10^(-9)个转导子/噬菌斑形成单位(PFU)。对于混合细菌群落,在进行的六个实验中有两个检测到了推定的转导子。通过用nptII探针进行菌落杂交以及用两组针对pQSR50的特异性引物进行PCR扩增,对这些推定的转导子进行了确认,并将其与本土抗生素抗性细菌分离。混合细菌群落中质粒转导的频率范围为1.58×10^(-8)至3.7×10^(-8)个转导子/PFU。使用数值模型获得的转导速率估计表明,坦帕湾河口每年可能发生多达1.3×10^(14)次转导事件。这项研究的结果表明,转导可能是海洋环境中水平基因转移的一种重要机制。