Cloning and characterization of glutamate dehydrogenase (GDH) from the gut of Haemonchus contortus
Parasitology - Tập 118 Số 3 - Trang 297-304 - 1999
Philip Skuce1, Elise M. Stewart2, W.D. Smith2, David P. Knox2
1Moredun Research Institute, International Research Centre, Penicuik, Midlothian, UK.
2Moredun Research Institute, International Research Centre, Pentlands Science Park, Bush Loan, Penicuik, Midlothian, EH26 0PZ, UK
Tóm tắt
Vaccination of lambs with the membrane-bound (S3) thiol-Sepharose
binding protein (TSBP) fraction derived from the
gut of Haemonchus contortus confers significant protection against
homologous challenge. The S3 TSBP peptide profile
is dominated by a major protein of ca. 60 kDa which is strongly
recognized by antisera from sheep demonstrably protected
following immunization with S3 TSBP. In an attempt to identify this protein,
sera from protected lambs were employed
to screen a λgt11 cDNA library of the adult parasite and resulted
in the isolation of numerous clones encoding a homologue
of the mitochondrial enzyme, glutamate dehydrogenase (GDH). GDH enzyme
activity was readily demonstrable in
S3 TSBP material and immunolocalization studies showed that the enzyme
was localized to the cytoplasm of the parasite's
gut. Furthermore, the enzyme appeared to be developmentally regulated,
with both GDH mRNA and protein expressed
almost exclusively during the blood-feeding parasitic stages.
