Recombinant vaccinia virus expression of Anaplasma marginale surface protein MSP-1a: effect of promoters, leader sequences and GPI anchor sequence on antibody response.
Anaplasma marginale surface protein MSP-1a was expressed by recombinant vaccinia viruses with different promoters and as hybrid proteins. Transcription of msp1 alpha with P11 late promoter resulted in more MSP-1a than with P7.5 early-late promoter; however, mice immunized with the recombinants had similar antibody titres. Recombinants expressing hybrid MSP-1a with either a murine leukaemia virus or a trypanosomal glycoprotein signal sequence did not enhance antibody responses and resulted in a diffuse intracellular distribution of MSP-1a which did not accumulate in the Golgi apparatus as was noted in the absence of these signal sequences. In contrast, antibody titres to MSP-1a in mice immunized with a recombinant virus expressing hybrid MSP-1a with a trypanosomal GPI anchor signal sequence were significantly increased over all other constructs.
Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman 99164-7040.
T C
TC
McGuire
E B
EB
Stephens
G H
GH
Palmer
T F
TF
McElwain
C A
CA
Lichtensteiger
S R
SR
Leib
A F
AF
Barbet
D000709:Anaplasma / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis* / Q000276:immunology; D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D002460:Cell Line; D002522:Chlorocebus aethiops; D004797:Enzyme-Linked Immunosorbent Assay; D005798:Genes, Bacterial; D017261:Glycosylphosphatidylinositols / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D017261:Glycosylphosphatidylinositols; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins