Introduction of bacteriophage Mu into bacteria of various genera and intergeneric gene transfer by RP4::Mu.

Abstract

The host range of coliphage Mu was greatly expanded to various genera of gram-negative bacteria by using the hybrid plasmid RP4::Mu cts, which is temperature sensitive and which confers resistance to ampicillin, kanamycin, and tetracycline. These drug resistance genes were transferred from Escherichia coli to members of the genera Klebsiella, Enterobacter, Citrobacter, Salmonella, Proteus, Erwinia, Serratia, Alcaligenes, Agrobacterium, Rhizobium, Pseudomonas, Acetobacter, and Bacillus. Mu phage was produced by thermal induction from the lysogens of all these drug-resistant bacteria except Bacillus. Mu phage and RP4 or the RP4: :Mu plasmid were used to create intergeneric recombinant strains by transfer of some genes, including the arylsulfatase gene, between Klebsiella aerogenes and E. coli. Thus, genetic analysis and intergeneric gene transfer are possible in these RP4: :Mu-sensitive bacteria.

Conjugation and heterologous gene expression in Gluconobacter oxydans ssp. suboxydans

“…To our knowledge, there has only been one reported attempt to conjugally transfer a broad host range plasmid to Acetobacter suboxydans (presently Gluconobacter suboxydans). Murooka et al [2] described the introduction of the P-group plasmid RP4::Mu into this strain; however, the conjugal transfer frequency for RP4 per se into this strain was not reported. In view of the industrial importance of Gluconobacter spp., it was decided to investigate further conjugal transfer of a series of broad host range plasmids into G. oxydans ssp.…”

“…This frequency was approximately 10 -6 exconjugants per recipient cell or 10 -5 per donor. 175 Murooka et al [2] did not report a transfer frequency for RP4.…”

Transfer of broad host range plasmids to the type I obligate methanotroph Methylomonas albus

“…Q and W have the ability to be stably maintained in a wide range of Gram-negative bacteria. Derivatives of these plasmids have been successfully used both as donors for transposon mutagenesis and as vectors for gene cloning [7][8][9]. Plasmids RP4 and R68.45 of the IncP group have been shown to transfer to Methylosinus trichosporium OB3b, a type 2 obligate methanotroph, but not to Methylococcus sp.…”

Transposition of Tn1831 to sym plasmids of Rhizobium leguminosarum and Rhizobium trifolii

“…When an R-factor in which the DNA of bacteriophage Mu is integrated is transferred from Escherichia coli to Rhizobium, it is not stably maintained in the new host [1,2]. Current informa-tion about the suicidal behaviour of these R::Mu plasmids in Rhizobium and Agrobacterium suggests that both a Mu function and a restriction mechanism are involved in preventing establishment of these plasmids [2][3][4]. This suicide character of R: :Mu plasmids offers a simple method to introduce transposons into Rhizobium.…”