Journal:

Article Title: Characterization of a WaaF (RfaF) Homolog Expressed by Haemophilus ducreyi

doi:

Figure Lengend Snippet: Bacterial strains and plasmids used in this study

Article Snippet: In these growth studies, H. ducreyi strains were grown at 33°C with slow shaking (90 rpm) in a water bath; growth was monitored by measurement of culture turbidity. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Description Reference or source Bacterial strains E. coli DH5α Host strain for cloning experiments 59 HB101 Host strain essential for propagating plasmids carrying mutated H. ducreyi DNA inserts used in electroporation 6 , 59 S. typhimurium SL3770 Expresses wild-type LPS 58 SL3789 rfaF ( waaF ) mutant; expresses a truncated LPS molecule which migrates rapidly in SDS-PAGE relative to SL3770 LPS 58 H. ducreyi 35000 Wild-type strain isolated in Winnipeg, Manitoba, Canada; LOS binds MAb 3E6 23 35000.7 Isogenic LOS mutant with a cat cartridge inserted into the rfaK ( lbgB ) gene; expresses a truncated LOS molecule that does not bind MAb 3E6 68 35000.252 Isogenic LOS mutant with a cat cartridge inserted into the Ppu 10I site within the gmhA gene; expresses a truncated LOS molecule that does not bind MAb 3E6 6 35000.10 Isogenic LOS mutant with an Ω-Cm cartridge inserted into the Msc I site within the waaF gene; expresses a truncated LOS molecule that does not bind MAb 3E6 This study Plasmids pUC19 Cloning vector; encodes Amp r 59 pHD1.3 pUC19 containing a 1.5-kb insert of H. ducreyi 35000 chromosomal DNA with an incomplete waaF ORF This study pBR322 Cloning vector; encodes Amp r , Tet r 59 pSLR1 pBR322 with a 5.8-kb H. ducreyi 35000 DNA insert containing the waaF gene This study pBB10 pSLR1 with an Ω-Cm cartridge inserted into the Msc I site within the waaF gene This study pCR2.1 Cloning vector; encodes Amp r Invitrogen pBB12 pCR2.1 with a 1.5-kb PCR-derived insert containing the H. ducreyi waaF gene This study pLS88 Cloning vector capable of replication in H. ducreyi ; encodes Kan r , Sm r , Sul r 15 pBB13 pLS88 with a 1.5-kb PCR-derived insert containing the H. ducreyi waaF gene This study Open in a separate window Bacterial strains and plasmids used in this study

Techniques: Plasmid Preparation, Clone Assay, Electroporation, Mutagenesis, Isolation

Journal:

Article Title: Characterization of a WaaF (RfaF) Homolog Expressed by Haemophilus ducreyi

doi:

Figure Lengend Snippet: Partial restriction map of the H. ducreyi 35000 chromosomal DNA insert in pSLR1 and related plasmids. The eight ORFs present in this insert are indicated together in boxes, with open arrows designating the proposed directions of transcription. Restriction sites in parentheses indicate vector cloning sites. The closed arrows indicate various oligonucleotide primers used in PCR. The cross-hatched bar beneath the waaF gene indicates the 328-bp probe used for Southern blot analysis (see Fig. ​Fig.4).4). The Ω-Cm cartridge was ligated into the MscI site of pSLR1 to construct pBB10. Plasmid pBB12 is pCR2.1 with a 1.5-kb PCR product containing the waaF gene and flanking DNA; the small boxes on the ends of this insert represent nucleotides involved in cloning it into the pCR2.1 vector. Plasmid pBB13 is pLS88 with the aforementioned 1.5-kb PCR product cloned into the EcoRI and SacI sites of this vector.

Article Snippet: In these growth studies, H. ducreyi strains were grown at 33°C with slow shaking (90 rpm) in a water bath; growth was monitored by measurement of culture turbidity. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Description Reference or source Bacterial strains E. coli DH5α Host strain for cloning experiments 59 HB101 Host strain essential for propagating plasmids carrying mutated H. ducreyi DNA inserts used in electroporation 6 , 59 S. typhimurium SL3770 Expresses wild-type LPS 58 SL3789 rfaF ( waaF ) mutant; expresses a truncated LPS molecule which migrates rapidly in SDS-PAGE relative to SL3770 LPS 58 H. ducreyi 35000 Wild-type strain isolated in Winnipeg, Manitoba, Canada; LOS binds MAb 3E6 23 35000.7 Isogenic LOS mutant with a cat cartridge inserted into the rfaK ( lbgB ) gene; expresses a truncated LOS molecule that does not bind MAb 3E6 68 35000.252 Isogenic LOS mutant with a cat cartridge inserted into the Ppu 10I site within the gmhA gene; expresses a truncated LOS molecule that does not bind MAb 3E6 6 35000.10 Isogenic LOS mutant with an Ω-Cm cartridge inserted into the Msc I site within the waaF gene; expresses a truncated LOS molecule that does not bind MAb 3E6 This study Plasmids pUC19 Cloning vector; encodes Amp r 59 pHD1.3 pUC19 containing a 1.5-kb insert of H. ducreyi 35000 chromosomal DNA with an incomplete waaF ORF This study pBR322 Cloning vector; encodes Amp r , Tet r 59 pSLR1 pBR322 with a 5.8-kb H. ducreyi 35000 DNA insert containing the waaF gene This study pBB10 pSLR1 with an Ω-Cm cartridge inserted into the Msc I site within the waaF gene This study pCR2.1 Cloning vector; encodes Amp r Invitrogen pBB12 pCR2.1 with a 1.5-kb PCR-derived insert containing the H. ducreyi waaF gene This study pLS88 Cloning vector capable of replication in H. ducreyi ; encodes Kan r , Sm r , Sul r 15 pBB13 pLS88 with a 1.5-kb PCR-derived insert containing the H. ducreyi waaF gene This study Open in a separate window Bacterial strains and plasmids used in this study

Techniques: Plasmid Preparation, Clone Assay, Southern Blot, Construct

Journal:

Article Title: Characterization of a WaaF (RfaF) Homolog Expressed by Haemophilus ducreyi

doi:

Figure Lengend Snippet: Comparison of the deduced amino acid sequences of the WaaF proteins from H. influenzae (20, 44), H. ducreyi 35000, S. typhimurium (65), and N. gonorrhoeae (53, 63) by using the Clustal-W alignment program in MacVector version 6. Dark shading indicates residues that are identical; light shading indicates residues that are similar.

Article Snippet: In these growth studies, H. ducreyi strains were grown at 33°C with slow shaking (90 rpm) in a water bath; growth was monitored by measurement of culture turbidity. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Description Reference or source Bacterial strains E. coli DH5α Host strain for cloning experiments 59 HB101 Host strain essential for propagating plasmids carrying mutated H. ducreyi DNA inserts used in electroporation 6 , 59 S. typhimurium SL3770 Expresses wild-type LPS 58 SL3789 rfaF ( waaF ) mutant; expresses a truncated LPS molecule which migrates rapidly in SDS-PAGE relative to SL3770 LPS 58 H. ducreyi 35000 Wild-type strain isolated in Winnipeg, Manitoba, Canada; LOS binds MAb 3E6 23 35000.7 Isogenic LOS mutant with a cat cartridge inserted into the rfaK ( lbgB ) gene; expresses a truncated LOS molecule that does not bind MAb 3E6 68 35000.252 Isogenic LOS mutant with a cat cartridge inserted into the Ppu 10I site within the gmhA gene; expresses a truncated LOS molecule that does not bind MAb 3E6 6 35000.10 Isogenic LOS mutant with an Ω-Cm cartridge inserted into the Msc I site within the waaF gene; expresses a truncated LOS molecule that does not bind MAb 3E6 This study Plasmids pUC19 Cloning vector; encodes Amp r 59 pHD1.3 pUC19 containing a 1.5-kb insert of H. ducreyi 35000 chromosomal DNA with an incomplete waaF ORF This study pBR322 Cloning vector; encodes Amp r , Tet r 59 pSLR1 pBR322 with a 5.8-kb H. ducreyi 35000 DNA insert containing the waaF gene This study pBB10 pSLR1 with an Ω-Cm cartridge inserted into the Msc I site within the waaF gene This study pCR2.1 Cloning vector; encodes Amp r Invitrogen pBB12 pCR2.1 with a 1.5-kb PCR-derived insert containing the H. ducreyi waaF gene This study pLS88 Cloning vector capable of replication in H. ducreyi ; encodes Kan r , Sm r , Sul r 15 pBB13 pLS88 with a 1.5-kb PCR-derived insert containing the H. ducreyi waaF gene This study Open in a separate window Bacterial strains and plasmids used in this study

Techniques: