|
KMID : 0613820010110040385
|
|
Journal of Life Science
2001 Volume.11 No. 4 p.385 ~ p.391
|
|
|
Expression of recombinant Bordetella pertussis filamentous hemagglutinin (FHA) antigen in Live Attenuated Salmonella typhimurium Vaccine Strain
|
|
Kang Ho-Young
|
|
|
Abstract
|
|
|
|
Filamentous hemagglutinin (FHA) is considered as an essential immunogenic component for incorporation into acellular vaccines against Bordetella pertussis, the causative agent of whooping cough. Classically, antipertussis vaccination has employed an intramuscular route. An alternative approach to stimulate mucosal and systemic immune responses is oral immunization with recombinant live vaccine carrier strains of Salmonella typhimurium. An attenuated live Salmonella vaccine sgrain(Deltacya Deltacrp) expressing recombinant FHA(rFHA) was developed. Stable expressionof rFHA was achieved by the use of balanced-lethal vector-host system. which employs an asd deletion in the host chromosome to impose in obligate requirement for diaminopimelic acid. The chromosomal Deltaasd mutation was complemented by a plasmid vector possessing the asd^{+} gene. A 3 kb DNA fragment encoding immuno dominant regionof FHA was subcloned in-frame downstream to the ATG translation initiation codon in the multicopy Asd^{+} pYA3341 vector to create pYA3457. Salmonella vaccine harboring pYA3457 expressed approximately 105kDa rFHA protein. The 100% maintenance of [YA3457 in vaccine strain was confirmed by stability examinations. Additionally, a recombinant plasmid pYA3458 was constructed to overpress His(8X)-tagged rFHA in Essherichia coli. His-tagged rFHA was purified from the E. coli strain harboring pYA3458 using Ni^{2+}-NTA affinity purification system.>^{2+}-NTA affinity purification system.
|
|
|
KEYWORD
|
|
|
Salmonella, Bordetella, Vaccine, immune response, pertussis
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|