Use of bacteriophage Mu to isolate deletions in the his-nif region of Klebsiella pneumoniae

Bachhuber, Mary; Brill, Winston J.; Howe, M M

doi:10.1128/jb.128.3.749-753.1976

Cited by 42 publications

(20 citation statements)

References 13 publications

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“…The bacteriophage Mu is a powerful genetic tool for isolating polar, nonreverting mutations due to prophage insertion and also for the generation of deletion mutations (19). Recently Mu has been used to generate deletions in the hisnif region of K. pneumoniae (4) and also in the plasnid pTM4010 (24). The plasnid pTM4010 is a derivative of pRD1, a recombinant R factor containing the K. pneumoniae his-nif region which no longer confers resistance to P1 and Mu (11,24).…”

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confidence: 99%

“…(Part of this work has been presented previously [T. MacNeil, G. P. Roberts (4) and for soft agar and minimal medium have been descibed (24). Solid media contained 15 g of agar (Difco Laboratories, Detroit, Mich.) per liter except N-free media, which contained 13 g of purified agar (Difco) per liter.…”

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confidence: 99%

“…Strains of K. pneumoniae and E. coli K-12 used in this work are listed in Table 1. UN727 and UN729 are Mu-sensitive derivatives of UN isolated as mutants resistant to a K. pneumoniae virulent phage (4). UN729 is moderately Mu sensitive and Plkc resistant, but Plkc hl sensitive.…”

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confidence: 99%

“…PlCMclrlOO (31) and Mu cts6i and Mu c25 have been described (1,18). Mu lysates were prepared as described previously (4).…”

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confidence: 99%

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Fine-structure mapping and complementation analysis of nif (nitrogen fixation) genes in Klebsiella pneumoniae

MacNeil¹,

MacNeil²,

Roberts³

et al. 1978

J Bacteriol

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158

110

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Four hundred and eighty-nine independent Nif-strains containing 260 point, 130 Mu-induced, and 99 deletion mutations in nif in the Klebsiella pneumoniae chromosome were isolated. Three hundred and ninety insertion and point mutations were mapped with Mu-induced deletions carried on 44 plasmids derived from pTM4010, a recombinant R factor containing the his-nif region of K. pneumoniae. The 99 chromosomal deletions in the nif region were mapped with 69 derivatives of pTM4010 carrying insertion and point mutations in nif. Complementation analysis between 84 derivatives of pTM4010 carrying nif mutations and Rec-derivatives of the 390 Nif-mutants identified 14 genes. The nif mutations were ordered into 49 deletion groups with a gene order of his... nifQBALFMVSNEKDHJ. Complementation analysis of Mu-induced, amber, frameshift, and deletion mutations indicates there are five polycistronic and two monocistronic operons: nifQ nifB, nifA nifL, nifF, nifM nifV nifS, nifN nifE, nifK nifD nifH, and nifJ. Transcription is from right to left in all polycistronic operons.

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confidence: 99%

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confidence: 99%

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confidence: 99%

“…PlCMclrlOO (31) and Mu cts6i and Mu c25 have been described (1,18). Mu lysates were prepared as described previously (4).…”

mentioning

confidence: 99%

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Fine-structure mapping and complementation analysis of nif (nitrogen fixation) genes in Klebsiella pneumoniae

MacNeil¹,

MacNeil²,

Roberts³

et al. 1978

J Bacteriol

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158

110

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“…Both phages naturally reside as prophages in HY and were removed from it by curing (STEIGER 1973). Similar changes in phage sensitivity are also known from other systems (BACHHUBER et al 1976). In the investigated example ink-34B no O-specific polysaccharides of the LPS were detectable.…”

Section: Discussionmentioning

confidence: 54%

Two genes in prophage y ofSerratia marcescens with functional counterparts in prophage Psi tested by Mu insertions as to order and homology

Steiger

Konhäuser

Alber

1999

J. Basic Microbiol.

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Two special genes carried in pairs by the native prophages y and ψ of Serratia marcescens HY, with functionally rather similar counterparts each, were assigned to restriction fragments and tested for homology. The y genes any and sky, as well as the ψ genes anp and skp, are specifically activated by infection of HY cells with κ phage. The κ genes exerting this effect are tay for y and tap for ψ. By means of tay and tap mutants, insertions of phage Mu DNA in the relevant parts of y and ψ prophage, respectively, could be discovered. These insertions and subsequent deletions of Mu were the basis of our studies. The use of Mu was made possible by the isolation of an HY variant giving appropriate plaques with Mu particles of the G(–)) type. In the case of another HY variant, Mu plaque formation depended on the presence of a host range mutation isolated by its ability to allow plaque formation on E. coli C, an indicator only for G(–) particles. Unexpectedly, this HY strain was an indicator of both G(–) and G(+) particles, but unfortunately had become adsorption resistant to y and ψ. As an accessory result, it provided evidence for a second restriction/modification system. In both cases the O‐specific poly‐saccharides were reduced. The main results of our paper concerning the two pairs of y and ψ genes are as follows: the orders any‐sky and anp‐skp correspond with each other in y and ψ; the sky gene, just as the skp gene, lies near one prophage end. However, despite the similarity in function and order, these genes are not homologous, in contrast to any and anp, which are at least partially homologous. The homologous regions of y and ψ amount to only about 0.5 kbp. Another observation was that bacteria with a Mu insertion near the skp end of y prophage were no longer cured of ψ when infected with κ tay‐1, in contrast to the efficient curing observed with an ordinary ψ prophage.

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Historical Perspective — Development of nif Genetics and Regulation in Klebsiella pneumoniae

Dixon

Genetics and Regulation of Nitrogen Fixation in Free-Living Bacteria

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Use of bacteriophage Mu to isolate deletions in the his-nif region of Klebsiella pneumoniae

Cited by 42 publications

References 13 publications

Fine-structure mapping and complementation analysis of nif (nitrogen fixation) genes in Klebsiella pneumoniae

Fine-structure mapping and complementation analysis of nif (nitrogen fixation) genes in Klebsiella pneumoniae

Two genes in prophage y ofSerratia marcescens with functional counterparts in prophage Psi tested by Mu insertions as to order and homology

Historical Perspective — Development of nif Genetics and Regulation in Klebsiella pneumoniae

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