Construction of intergeneric hybrids using bacteriophage P1CM: transfer of the Klebsiella aerogenes ribitol dehydrogenase gene to Escherichia coli

Abstract

Study of many of the interesting properties of K. aerogenes is limited by the lack of a well characterized genetic system for this organism. Investigations of the evolution of the enzyme ribitol dehydrogenase (EC 1.1.1.56) in K. aerogenes would be greatly facilitated by the availability of such a system, and two approaches to developing one are reported here. The authors isolated mutants sensitive to the coliphage P1, which will efficiently transduce genetic markers between such sensitive strains and which will thus make detailed mapping studies possible. Derivatives of K. aerogenes lysogenic for P1 can be readily isolated by using the specialized transducing particle P1CMclr100. Bacteria lysogenic for this phage are chloramphenicol resistant and temperature sensitive. Phage particles produced by temperature induction of such lysogens can be used to transfer K. aerogenes genes to the natural host of P1 phage, Escherichia coli. This method was used to prepare derivatives of E. coli K 12 carrying the K. aerogenes conferring the ability to metabolize the pentitols ribitol and D arabitol. It was shown that these E. coli K. aerogenes hybrids synthesize a ribitol dehydrogenase with the properties of the K. aerogenes enzyme and the position of the transferred gene on the E. coli chromosome. The ramifications of this methodoloy are discussed.