Fundamental study of expression-library immunization against Taenia taeniaeformis in rats.
Expression-library immunization has been proposed as an effective means to screen a large number of genes of the pathogen as candidate protective molecules. In this study, we examined the efficacy of expression-library immunization using a T. taeniaeformis rat model system. Total RNAs were isolated from the last 15 segments of adult T. taeniaeformis and poly A RNA was purified. cDNA library was produced using SuperScript Plasmid System, which contains a mammalian expression vector, pCMV*SPORT6. From about 3,500 clones examined, more than 800 clones were found to contain DNA fragments. About 200 clones were sequenced and the homology search was carried out. The blast search revealed that 29% of the expression genes were mitochondrial genes (rRNA; 17%, protein; 12%). Nuclear rRNA genes (10%), nuclear protein (9%) and genes from Escherichia coli were also detected. Forty-two percent of sequences did not show a significant similarity to any genes deposited in the public database. Rats were immunized with expression-library and injected orally with 1,000 T. taeniaeformis eggs. However the protective effect of expression-library vaccine was not confirmed.
Okamoto
Munehiro
M
Department of Laboratory Animal Science, School of Veterinary Medicaine, Faculty of Agriculture, Totori University, Japan. mokamoto@muses.tottori-u.ac.jp
Matsumoto
Kanako
K
Shibui
Akiko
A
Shibahara
Toshiyuki
T
Ito
Akira
A
D000818:Animals; D000909:Antibodies, Helminth; D005260:Female; D015870:Gene Expression; D007114:Immunization; D015138:RNA, Nuclear; D051381:Rats; D013621:Taenia / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D013622:Taeniasis / Q000276:immunology / Q000469:parasitology* / Q000517:prevention & control; D019444:Vaccines, DNA / Q000494:pharmacology
D000909:Antibodies, Helminth; D015138:RNA, Nuclear; D019444:Vaccines, DNA