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Table 3 PCR/RFLP typing of fliC genes in E. coli H4 strains with restriction enzymes HhaI, HpaII and MboI

From: Genetical and functional investigation of fliC genes encoding flagellar serotype H4 in wildtype strains of Escherichia coli and in a laboratory E. coli K-12 strain expressing flagellar antigen type H48

 

DNA fragments (bp) obtained by enzymatic digestion of the 953 bp PCR product obtained with primers fliC-1 and fliC-2

fliC-genotype (prototype strain)

HhaI

HpaII

MboI

fliC-H4 (U9-41)a

362, 304, 104, 66, 50, 29d, 20d, 16d 2d

296, 240, 225, 159, 33d

429, 313, 182, 29d

fliC-H4 (P7d)b

362, 304, 104, 66, 50, 29d, 20d, 16d,2d

296, 273, 225, 159

429, 313, 182, 29d

fliC-H4 (P12b)c

362, 304, 104, 66, 50, 29d, 20d, 16d,2d

296, 273, 225, 159

495, 458

  1. a) PCR/RFLP patterns found in strains U9-41, U1-41, E1541-68, C107-74 and in 82 E. coli strains which reacted with H4 antisera. The serotypes of all 86 strains are: O2:K1 (4 strains), O5 (2), O7 (1), O8 (1), O13 (1), O15 (2), O20 (2), O50 (1), O60 (1), O77 (1), O78 (1), O99 (3), O111 (9), O113 (32), O114 (5), O117 (1), O119 (1), O141 (3), O154 (1), O176 (1), O181 (1), O-untypable (9), O-rough (3). Forty-two of the strains belonging to O-groups O60, O113, O114, O141 as well as O-rough and O-untypable strains produced Shiga-toxins.
  2. b) RFLP patterns found with strain P7d (O68:H4)
  3. c) RFLP-patterns found with strain P12b (O15:H17)
  4. d) fragments smaller than 50 bp are not detectable on 2% agarose gels (Fig. 3). Exact fragment sizes were calculated on the basis of nucleotide sequence analysis for each of the restriction enzymes used