Bremer E, Silhavy T J, Weinstock G M
Department of Biology, University of Konstanz, F.R.G.
Gene. 1988 Nov 15;71(1):177-86. doi: 10.1016/0378-1119(88)90089-3.
Lambda placMu phages are derivatives of bacteriophage lambda that use the transposition machinery of phage Mu to insert into chromosomal and cloned genes. When inserted in the proper fashion, these phages yield stable fusions to the Escherichia coli lac operon in a single step. We have determined the amount of DNA from the c end of phage Mu present in one of these phages, lambda placMu3, and have shown that this phage carries a 3137-bp fragment of Mu DNA. This DNA segment carries the Mu c-end attachment site and encodes the Mu genes cts62, ner+, and gene A lacking 179 bp at its 3' end (A'). The product of this truncated gene A' retains transposase activity and is sufficient for the transposition of lambda placMu. This was demonstrated by showing that lambda placMu derivatives carrying the A am1093 mutation in the A' gene are unable to transpose by themselves in a Su- strain, but their transposition can be triggered by coinfection with lambda pMu507(A+ B+). We have constructed several new lambda placMu phages that carry the A' am1093 gene and the kan gene, which confers resistance to kanamycin. Chromosomal insertions of these new phages are even more stable than those of the previously reported lambda placMu phages, which makes them useful tools for genetic analysis.
λplacMu噬菌体是噬菌体λ的衍生物,它们利用噬菌体Mu的转座机制插入染色体基因和克隆基因。当以适当方式插入时,这些噬菌体可一步产生与大肠杆菌乳糖操纵子的稳定融合。我们已确定这些噬菌体之一λplacMu3中存在的来自噬菌体Mu c末端的DNA量,并表明该噬菌体携带一段3137 bp的Mu DNA片段。该DNA片段携带Mu c末端附着位点,并编码Mu基因cts62、ner+以及在其3'末端缺失179 bp的基因A(A')。这个截短的基因A'的产物保留转座酶活性,并且对于λplacMu的转座是足够的。这通过以下方式得到证明:携带A'基因中A am1093突变的λplacMu衍生物在Su-菌株中自身无法转座,但它们的转座可由与λpMu507(A + B +)共感染触发。我们构建了几种携带A' am1093基因和赋予卡那霉素抗性的卡那基因的新λplacMu噬菌体。这些新噬菌体的染色体插入比先前报道的λplacMu噬菌体的插入更稳定,这使它们成为遗传分析的有用工具。