However, no putative integrase or mobility-associated genes were identified. Open reading frame (ORF) and BLAST analyses were performed on the KpGI-5 sequence (Figure 1 and Table 1). The 2.7 kb segment mapping
KU-60019 datasheet to the right arm of KpGI-5 was 90% identical to a region immediately downstream of met56 in K. pneumoniae Kp342 and was predicted to encode two hypothetical BAY 63-2521 datasheet proteins (Orf14 and Orf15), a metallo-beta-lactamase family protein (Orf16) and a putative GCN5-related N-acetyltransferase (Orf13). The nucleotide sequence of a 3.4 kb central region did not match any GenBank entries and coded for three novel proteins; Orf10 and Orf11 exhibited weak matches to putative regulatory proteins from the bacteria Stigmatella aurantiaca DW4/3-1 and Serratia odorifera DSM 4582, respectively. Orf10 also possessed a match to the pfam domain Trans_reg_C (PF00486) which has been
see more implicated in DNA binding, further suggesting a role for Orf10 in regulation. Figure 1 Features of the novel KpGI-5 genomic island in K. pneumoniae KR116. (A) Genetic organisation of KpGI-5 shown lying between the species-conserved upstream flank (UF) and downstream flank (DF) sequences. The eight putative fimbrial genes are labelled fim2A–fim2K. Closest BLASTP similarities for these and other predicted KpGI-5-encoded proteins are described in Table 1. KpGI-5 segments indicated by double arrows map to G + C % transitions as indicated by G + C profile. The thin horizontal
lines on the G + C % graph represent the average G + C content of the K. pneumoniae MGH78578 genome (57.4%) and the entire KpGI-5 island (44.0%). The 20.8% and 65.0% G + C content lines correspond to the minimum and maximum G + C % calculated over an 80 bp window, respectively. (B) Alignment of the tRNA-proximal (DRP) and tRNA-distal (DRD) 46 bp direct repeat (DR) sequences associated with KpGI-5. DRP comprises the 3’ end of met56. Table 1 BLASTP homologs of proteins predicted GPCR & G Protein inhibitor to be encoded by KpGI-5 Gene name Coding region (bp) Protein size (aaa) Percentage identity (aaa) Organism Possible function [GenBank Number] E value met56 180..255 (76) / 100% (note: BLASTN) K. pneumoniae MGH78578 Methionine tRNA [KPN_03476] / fim2K 1385..528 (858) 285 60% (165/276) C. koseri ATCC BAA-895 Putative EAL domain protein [ABV14791.1] 1e-94 fim2H 2440..1514 (927) 308 62% (190/308) K. pneumoniae sp15 Fimbrial adhesin (FimH) [ACL13802.1] 1e-101 fim2G 2961..2458 (504) 167 72% (120/167) C. koseri ATCC BAA-895 Minor fimbrial subunit (FimG) [ABV14789.1] 2e-65 fim2F 3501..2974 (528) 175 79% (138/175) C. koseri ATCC BAA-895 Minor fimbrial subunit (FimF) [ABV14788.1] 1e-73 fim2D 6073..3515 (2559) 852 82% (689/838) C. koseri ATCC BAA-895 Outer membrane usher protein (FimD) [ABV14787.1] 0.0 fim2C 6858..6229 (630) 209 92% (190/207) K. variicola At-22 Fimbrial chaperone protein (FimC) [ADC56706.1] 2e-107 fim2I 7519..6989 (531) 176 82% (139/170) C.