Duzhiĭ D E, Zavil'gel'skiĭ G B
Mol Gen Mikrobiol Virusol. 1994 May-Jun(3):36-8.
The bacteriophages lambda:lux and lambda:luxAB have been constructed by ligation of phage arms generated by BamHI or SalGI restriction endonucleases digestion of EMBL4 to BamHI digested plasmid pF1 lux+ or to SalGI digested plasmid pF2 lambda:luxA+B+. Cells of Escherichia coli prototrophic strain Cs were infected with lambda:lux or lambda:luxAB and intensity of bioluminiscence of the samples registered at different time intervals determined. The signal of bioluminiscence was first detected 15 min after infection and its level increased exponentially thereafter demonstrating replication of the lambda:lux bacteriophages. We have used the recombinant lambda:luxAB bacteriophage to detect the enteric indicator bacteria without enrichment in 15 min, provided that they are present at levels higher than 10(4).
噬菌体λ:lux和λ:luxAB是通过将EMBL4经BamHI或SalGI限制性内切酶消化产生的噬菌体臂与经BamHI消化的质粒pF1 lux⁺或经SalGI消化的质粒pF2 λ:luxA⁺B⁺连接构建而成。用λ:lux或λ:luxAB感染大肠杆菌原养型菌株Cs的细胞,并在不同时间间隔测定样品的生物发光强度。感染后15分钟首次检测到生物发光信号,此后其水平呈指数增长,表明λ:lux噬菌体在复制。我们使用重组λ:luxAB噬菌体在15分钟内无需富集即可检测肠道指示菌,前提是它们的存在水平高于10⁴。