Table 1 Strains, plasmids and primers used in this study
Strains, plasmids and primers | Description | Source |
|---|---|---|
Strains | ||
 E. coli DH5α | F− Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rk−, mk+) phoA supE44 λ− thi −1 gyrA96 relA1 | Invitrogen |
 E. coli BL21 DE3 | F−ompTgaldcmlonhsdSB(rB-mB-)λ(DE3[lacIlacUV5-T7gene1ind1sam7nin5], expression host | Stratagene |
 Nontypeable H. influenza ATCC 49766 | The source of gene for protein E | Pasteur Institute of Iran |
Plasmids | ||
 pET-26b | T7promoter, an N-terminal pelB signal sequence for potential periplasmic localization, plus optional C-terminal His Tag sequence, kanamycin resistance | Novagen |
 pET-21b | T7promoter, an N-terminal T7 Tag sequence plus an optional C-terminal His Tag sequence, ampicillin resistance | Novagen |
 pGEM-JR | The source of gene for Lpp-OmpA | Bioneer |
 p21PE | Expression vector; Ampr | This study |
 pPE22-160 | Expression vector; Kanr | This study |
 pLpp-OmpA | Expression vector; Kanr | This study |
 pNon-OmpA | Expression vector; Kanr | This study |
 pLpp-OmpA-PE | Expression vector; Kanr | This study |
 pNon-OmpA-PE | Expression vector; Kanr | This study |
Primers | Sequence (5′–3′) | Restriction enzymes |
|---|---|---|
EcoRIpE22 | GTCAGAATTCAAAGGCTAAACAAAATGATGT | EcoRI |
HindIIIpE22 | GATCAAGCTTTTTTTTATCAACTGAAAATGC | HindIII |
NcoIpE22 | CATGCCATGGATAAGGCTAAACAAAATGATGTG | NcoI |
PENdeI | GGGCATCCATATGAAAAAAATTATTTTAACA | NdeI |
NonOmpAF | CCCATATGAAAGCTACTAAACTGGTACTGGGCAACCCGTATGTTGGCTTTGAAATGGG | NdeI |
NonOmpAR | CGGAATTCGCTCCCGGAATGCCGTTGTCCGGACGAGTGCC | EcoRI |