Expression of recombinant antigens in Escherichia coli: application on immunochemical studies of Schistosoma mansoni tegumental antigens.
Sm15 and Sm13 are recognized by antibodies from mice protectively vaccinated with tegumental membranes, suggesting a potential role in protective immunity. In order to raise antibodies for immunochemical investigations, the genes for these antigens were expressed in pGEX and pMal vectors so that comparisons could be made among different expression systems and different genes. The fusion proteins corresponding to several parts of the gene for the precursor of Sm15 failed in producing antibodies recognizing the parasite counterpart. On the other hand, antibodies raised against Sm13 MBP-fusion proteins recognized the 13 kDa tegumental protein. Thus the peculiarities of the gene of interest are important and the choice of the expression system must sometimes be decided on an empirical basis.
Abath
F G
FG
Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães-FIOCRUZ, Recife, PE, Brasil.
Xavier
E M
EM
Silva
J D
JD
Morais Júnior
M A
MA
Montenegro
S M
SM
D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D018396:Mucin-1 / Q000276:immunology; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012550:Schistosoma mansoni / Q000276:immunology*
D000947:Antigens, Helminth; D018396:Mucin-1; D011993:Recombinant Fusion Proteins