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Klebsiella pneumoniae subsp. pneumoniae-bacteriophage combination from the caecal effluent of a healthy woman.

Hoyles L, Murphy J, Neve H, Heller KJ, Turton JF, Mahony J, Sanderson JD, Hudspith B, Gibson GR, McCartney AL, van Sinderen D - PeerJ (2015)

Bottom Line: This Siphoviridae phage presents a rosette-like tail tip and exhibits depolymerase activity, as demonstrated by the formation of plaque-surrounding haloes that increased in size over the course of incubation.The genome of KLPN1 was determined to be 49,037 bp (50.53 %GC) in length, encompassing 73 predicted ORFs, of which 23 represented genes associated with structure, host recognition, packaging, DNA replication and cell lysis.On the basis of sequence analyses, phages KLPN1 (GenBank: KR262148) and 1513 (a member of the family Siphoviridae, GenBank: KP658157) were found to be two new members of the genus "Kp36likevirus."

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Microbiology, University College Cork , Cork , Ireland ; Department of Biomedical Sciences, University of Westminster , London , United Kingdom.

ABSTRACT
A sample of caecal effluent was obtained from a female patient who had undergone a routine colonoscopic examination. Bacteria were isolated anaerobically from the sample, and screened against the remaining filtered caecal effluent in an attempt to isolate bacteriophages (phages). A lytic phage, named KLPN1, was isolated on a strain identified as Klebsiella pneumoniae subsp. pneumoniae (capsular type K2, rmpA (+)). This Siphoviridae phage presents a rosette-like tail tip and exhibits depolymerase activity, as demonstrated by the formation of plaque-surrounding haloes that increased in size over the course of incubation. When screened against a panel of clinical isolates of K. pneumoniae subsp. pneumoniae, phage KLPN1 was shown to infect and lyse capsular type K2 strains, though it did not exhibit depolymerase activity on such hosts. The genome of KLPN1 was determined to be 49,037 bp (50.53 %GC) in length, encompassing 73 predicted ORFs, of which 23 represented genes associated with structure, host recognition, packaging, DNA replication and cell lysis. On the basis of sequence analyses, phages KLPN1 (GenBank: KR262148) and 1513 (a member of the family Siphoviridae, GenBank: KP658157) were found to be two new members of the genus "Kp36likevirus."

No MeSH data available.


Related in: MedlinePlus

Transmission electron micrographs of phage KLPN1.The phage belongs to the family Siphoviridae, and has an unusual base-plate structure, resembling a rosette. The arrows indicate the central tail fibre surrounded by three flexible spherical base-plate structures.
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fig-2: Transmission electron micrographs of phage KLPN1.The phage belongs to the family Siphoviridae, and has an unusual base-plate structure, resembling a rosette. The arrows indicate the central tail fibre surrounded by three flexible spherical base-plate structures.

Mentions: Phage KLPN1 is chloroform-resistant. In addition, it displayed stability to prolonged storage in TSB at 4 °C: after 6 and 18 months’ storage, titres for the phage were still 1010 pfu/ml, comparable with the original stock. Transmission electron microscopy revealed the isometric-headed phage to possess a capsid of ∼62.7 ± 2.3 nm (n = 30) in diameter with a long non-contractile tail of ∼164.4 ± 3.0 nm (n = 29), thus indicating that this phage is a member of the family Siphoviridae (Fig. 2). Notably, the base-plate structure was unusual with a distinct central tail fibre [length 33.4 ± 1.7 nm (n = 43)] with apparently three elongated spherical structures [length, 13.2 ± 1.3 nm (n = 43); width, 6.9 ± 0.7 nm (n = 48)] loosely associated with the central regions of the tail fibre. This unique base-plate structures resembles a rosette with three leaves, and can be observed in all five micrographs shown in Fig. 2. A similar structure has been described for Rtp, a lytic phage of Escherichia coli (Wietzorrek et al., 2006), with which KLPN1 shares some sequence identity (discussed below). It is also visible in electron micrographs of phage ϕ28, which infects K. pneumoniae capsular type K28 (Geyer et al., 1983).


Klebsiella pneumoniae subsp. pneumoniae-bacteriophage combination from the caecal effluent of a healthy woman.

Hoyles L, Murphy J, Neve H, Heller KJ, Turton JF, Mahony J, Sanderson JD, Hudspith B, Gibson GR, McCartney AL, van Sinderen D - PeerJ (2015)

Transmission electron micrographs of phage KLPN1.The phage belongs to the family Siphoviridae, and has an unusual base-plate structure, resembling a rosette. The arrows indicate the central tail fibre surrounded by three flexible spherical base-plate structures.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4525690&req=5

fig-2: Transmission electron micrographs of phage KLPN1.The phage belongs to the family Siphoviridae, and has an unusual base-plate structure, resembling a rosette. The arrows indicate the central tail fibre surrounded by three flexible spherical base-plate structures.
Mentions: Phage KLPN1 is chloroform-resistant. In addition, it displayed stability to prolonged storage in TSB at 4 °C: after 6 and 18 months’ storage, titres for the phage were still 1010 pfu/ml, comparable with the original stock. Transmission electron microscopy revealed the isometric-headed phage to possess a capsid of ∼62.7 ± 2.3 nm (n = 30) in diameter with a long non-contractile tail of ∼164.4 ± 3.0 nm (n = 29), thus indicating that this phage is a member of the family Siphoviridae (Fig. 2). Notably, the base-plate structure was unusual with a distinct central tail fibre [length 33.4 ± 1.7 nm (n = 43)] with apparently three elongated spherical structures [length, 13.2 ± 1.3 nm (n = 43); width, 6.9 ± 0.7 nm (n = 48)] loosely associated with the central regions of the tail fibre. This unique base-plate structures resembles a rosette with three leaves, and can be observed in all five micrographs shown in Fig. 2. A similar structure has been described for Rtp, a lytic phage of Escherichia coli (Wietzorrek et al., 2006), with which KLPN1 shares some sequence identity (discussed below). It is also visible in electron micrographs of phage ϕ28, which infects K. pneumoniae capsular type K28 (Geyer et al., 1983).

Bottom Line: This Siphoviridae phage presents a rosette-like tail tip and exhibits depolymerase activity, as demonstrated by the formation of plaque-surrounding haloes that increased in size over the course of incubation.The genome of KLPN1 was determined to be 49,037 bp (50.53 %GC) in length, encompassing 73 predicted ORFs, of which 23 represented genes associated with structure, host recognition, packaging, DNA replication and cell lysis.On the basis of sequence analyses, phages KLPN1 (GenBank: KR262148) and 1513 (a member of the family Siphoviridae, GenBank: KP658157) were found to be two new members of the genus "Kp36likevirus."

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Microbiology, University College Cork , Cork , Ireland ; Department of Biomedical Sciences, University of Westminster , London , United Kingdom.

ABSTRACT
A sample of caecal effluent was obtained from a female patient who had undergone a routine colonoscopic examination. Bacteria were isolated anaerobically from the sample, and screened against the remaining filtered caecal effluent in an attempt to isolate bacteriophages (phages). A lytic phage, named KLPN1, was isolated on a strain identified as Klebsiella pneumoniae subsp. pneumoniae (capsular type K2, rmpA (+)). This Siphoviridae phage presents a rosette-like tail tip and exhibits depolymerase activity, as demonstrated by the formation of plaque-surrounding haloes that increased in size over the course of incubation. When screened against a panel of clinical isolates of K. pneumoniae subsp. pneumoniae, phage KLPN1 was shown to infect and lyse capsular type K2 strains, though it did not exhibit depolymerase activity on such hosts. The genome of KLPN1 was determined to be 49,037 bp (50.53 %GC) in length, encompassing 73 predicted ORFs, of which 23 represented genes associated with structure, host recognition, packaging, DNA replication and cell lysis. On the basis of sequence analyses, phages KLPN1 (GenBank: KR262148) and 1513 (a member of the family Siphoviridae, GenBank: KP658157) were found to be two new members of the genus "Kp36likevirus."

No MeSH data available.


Related in: MedlinePlus