Čeština
EnglishProduction of Polyclonal Antiobies to a Recombinant Potato Mop-top Virus Non-structural Triple Gene Block Protein l
Čeřovská, Noemi; Filigarová, Marie; Pečenková, Tamara
JOURNAL OF PHYTOPATHOLOGY Phytopathologische Zeitschrift 154: 422-427, 2006
Keywords: Potato mop-top virus; recombinant protein; triple gene block; polyclonal antibodies
Abstract: The gene encoding the triple gene protein 1 (TGBp1) of Potato mop-top protein (PMTV) was cloned into expression vector pQE32 tagging the protein with 6xHis on the N-terminus. When the gene was enshortened on its 3´-end by two different restriction digestions, efficient and high yield bacterial expression was achieved in both cases, as shown by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PHAGE) and Western blot. One of these two constructs was used for raising rabbit polyclonal antibodies. The obtained sera and antibodies were tested for the detection of PMTV in laboratory host Nicotiana bethamiana and natural host Solanum tuberosum by enzyme-linked immunosorbent assay (ELISA) as well as by Western blota. The obtainad antisera are more suitable for Western blot analysis of infected plants than for ELSA.
DOI:
IEB authors: Noemi Čeřovská, Tamara Pečenková
JOURNAL OF PHYTOPATHOLOGY Phytopathologische Zeitschrift 154: 422-427, 2006
Keywords: Potato mop-top virus; recombinant protein; triple gene block; polyclonal antibodies
Abstract: The gene encoding the triple gene protein 1 (TGBp1) of Potato mop-top protein (PMTV) was cloned into expression vector pQE32 tagging the protein with 6xHis on the N-terminus. When the gene was enshortened on its 3´-end by two different restriction digestions, efficient and high yield bacterial expression was achieved in both cases, as shown by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PHAGE) and Western blot. One of these two constructs was used for raising rabbit polyclonal antibodies. The obtained sera and antibodies were tested for the detection of PMTV in laboratory host Nicotiana bethamiana and natural host Solanum tuberosum by enzyme-linked immunosorbent assay (ELISA) as well as by Western blota. The obtainad antisera are more suitable for Western blot analysis of infected plants than for ELSA.
DOI:
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