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Expression of Recombinant Potato leafroll virus Structural and Non‐structural Proteins for Antibody Production
Author(s) -
Plchova Helena,
Moravec Tomas,
Dedic Petr,
Cerovska Noemi
Publication year - 2011
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/j.1439-0434.2010.01740.x
Subject(s) - potato leafroll virus , biology , recombinant dna , virology , vector (molecular biology) , microbiology and biotechnology , escherichia coli , expression vector , capsid , virus , plant virus , gene , biochemistry
Vector pMPM‐A4Ω and vectors pQE‐30 and pET‐45b(+) containing the 6x His‐tag sequence were used for expression of Potato leafroll virus (PLRV) structural and non‐structural proteins in Escherichia coli . Coat protein (CP) and RNA‐dependent RNA polymerase (RdRp)–fragments RdRp 43‐616 and RdRp 304‐537 were chosen for expression. A high level of CP and RdRp 304‐537 was obtained only in an expression system using pET‐45b(+) vector and E. coli Rosetta‐gami 2(DE3) cells. After purification, the His‐tagged PLRV proteins were used for immunization of rabbits.