Expression of liver fluke antigens by mouse cells |
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| Authors: | P L Beardsell M J Howell |
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| Institution: | 1. Institute of Parasitic Diseases, Korea Association of Health Promotion, Seoul 07649, Republic of Korea;2. Department of Tropical Medicine and Parasitology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea;1. School of Biological Sciences, Queen''s University Belfast, Belfast, United Kingdom;2. The School of Life Sciences, University of Technology Sydney (UTS), Ultimo, Sydney, NSW, Australia;3. Department of Basic Pathology, Federal University of Parana, Curitiba, Brazil;4. School of Biotechnology, Dublin City University, Dublin, Republic of Ireland |
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| Abstract: | Fasciola hepatica cDNA carried on bacterial plasmids was used in conjunction with marker plasmid DNA to co-transform mouse tissue culture cells using the calcium phosphate procedure. Two systems were used: mouse L cells lacking thymidine kinase activity (Ltk−) in conjunction with plasmids pFH4 or pFH1 (carrying parasite DNA) and pHSV-106 (carrying the thymidine kinase gene from herpes simplex virus); and C127 mouse cells with the plasmids pFH4 or pFH1 and the plasmid pBPV-MMTneo(342-12) which carries the bovine papilloma virus genome and, as a selective marker, a gene conferring resistance to the antibiotic geneticin. Both procedures gave rise to transformants which expressed liver fluke antigens: these were detected by a fluorescent antibody test (incorporating flow cytometry) using fluke-infected sheep serum as first antibody. Stability of antigen expression characterised C 127 derived transformants. Ltk− transformants ceased expression within a few weeks. |
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