TABLE 2.
Description of mutations in C. sakazakii ES5 that result in reduced biofilm on polystyrene
| Functional group | Mutant(s)a | Locus tag (gene ID no.)b | Annotationc |
Reference(s) | Phenotype |
|||||
|---|---|---|---|---|---|---|---|---|---|---|
| Gene | Description (organism) | Accession no. | No. of positive amino acids/total no. (%) | Biofilm in CV assay (% of wild type, mean ± SD)g | CR agar | Flagella (EM) | ||||
| Cellulose biosynthesis | BF3, BF10 | ESA_04207 (5550384) | bcsC (yhjL) | Cellulose synthase subunit BcsC (Cronobacter sakazakii BAA-894)b | YP_001440 223.1 | 1,167/1,167 (100) | 15, 38, 52 | 33.6 ± 25.4 | sawh | NDk |
| BF6 | ESA_04199 (5550446) | bcsG (yhjU) | Conserved hypothetical protein (Cronobacter sakazakii)d | CAM32322.1 | 557/561 (99) | 15, 38, 52 | 20.5 ± 8.9 | saw | ND | |
| BF12 | ESA_04204 (5550385) | bcsA (yhjO) | Cellulose synthase catalytic subunit (Cronobacter sakazakii BAA-894)b | YP_001440 220.1 | 872/872 (100) | 15, 38, 52 | 15.5 ± 5.9 | saw | ND | |
| Flagellar structure or biosynthesis | BF2 | ESA_01356 (5549172) | flhE | Flagellar protein FlhE (Cronobacter turicensis) | CBA31743.1 | 118/119 (99) | 43 | 19.7 ± 6.3 | pdari | Like wild type |
| BF5 | ESA_01287 (5549196) | fliD | Flagellar hook-associated 2 domain protein 2 (Cronobacter turicensis) | CBA31883.1 | 471/476 (98) | 29, 48 | 9.3 ± 5.1 | pdar | Shorter, brittle | |
| BF13 | ESA_02266 (5551730) | flgJ | Peptidoglycan hydrolase (Cronobacter sakazakii BAA-894)b | YP_001438 351.1 | 321/321 (100) | 29 | 77.9 ± 6.4 | pdar | Absent | |
| Basic cellular processes | ND | |||||||||
| Cell division | BF1 | ESA_02449 (5549447) | ftsK | DNA translocase FtsK (Cronobacter turicensis) | CBA29614.1 | 1,264/1,364 (92) | 6 | 8.3 ± 2.6 | pdar | ND |
| Energy production | BF8 | ESA_02873 (5548784) | cyoD | Cytochrome o ubiquinol oxidase protein CyoD (Cronobacter turicensis) | CBA28615.1 | 109/109 (100) | 39 | 19.3 ± 6.1 | pdar | ND |
| BF9 | ESA_00861 (5550827) | alsS (ilvB) | Acetolactate synthase (Cronobacter sakazakii BAA-894)b | YP_001436 968.1 | 559/559 (100) | 36 | 16.5 ± 3.4 | pdar | ND | |
| Possibly involved in virulence | BF17 | ESA_pESA3p05536 (5552640) | mgtB | Magnesium-transporting ATPase, P-type 1 (Cronobacter turicensis) | CBA34629.1 | 853/904 (94) | 41 | 22.5 ± 9.1 | pdar | ND |
| Unknown function | BF4 | ESA_04103 (5551795) | NAe | Hypothetical protein Ent638_0111 (Enterobacter sp. 638) | ABP58801.1 | 361/425 (84) | NA | 66.9 ± 20.1 | pdar | Like wild type |
| BF11, BF15 | ESA_00282 (5549918) | NA | Hypothetical protein (Cronobacter turicensis) | CBA33797.1 | 164/166 (98) | NA | 72.8 ± 21.3 | pdarsej | Like wild type | |
| BF14 | ESA_00281 (5549972) | NA | Hypothetical protein (Cronobacter turicensis) | CBA33799.1.1 | 132/133 (99) | NA | 60.5 ± 36.9 | pdarse | Like wild type | |
| BF16 | NA | NA | Similar to Escherichia coli plasmids pMG828-5, p6148, and pE2348f | DQ995355.1, EU580136.1, and FM180570.1 | NA | NA | 34.3 ± 24.0 | pdar | ND | |
Transposon-containing derivative(s) of C. sakazakii ES5.
Based on the genome of C. sakazakii BAA-894 (http://www.ncbi.nlm.nih.gov/sites/entrez?db=genomeprj&cmd=Retrieve&dopt=Overview&list_uids=12720).
Based on a BlastP search with BAA-894 protein sequences of the NCBI nonredundant protein sequences on 15 January 2010.
C. sakazakii ES5 sequence from BAC clone (15).
NA, not applicable.
Based on BlastN search with the sequence obtained for ES5 mutant of the nucleotide collection (nr/nt) on 15 January 2010.
The value for the wild-type biofilm in the CV assay was 100% ± 9.2%.
saw, smooth and white (38).
pdar, pink, dry, and rough (38).
pdarse, pink, dry, and rough with smooth edge (this study).
ND, not determined.