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Nematollahi, Sevil,Sokhandan-Bashir, Nemat,Rakhshandehroo, Farshad,Zamanizadeh, Hamid Reza The Korean Society of Plant Pathology 2012 Plant Pathology Journal Vol.28 No.4
Molecular characterization of Cucumber mosaic virus (CMV) was done by using samples from tomato and cucurbitaceous plants collected from different locations in the northwest region of Iran. After screening by enzyme-linked immunosorbent assay, 91 CMV-infected samples were identified. Biological properties of eight representative isolates were compared with each other revealing two distinct phenotypes on squash and tomato plants. Phylogenetic analyses based on nucleotide sequences of the coat protein (CP), movement protein (MP) and 2b of the new isolates, together with that of previously reported isolates, led to the placement of the Iranian isolates in subgroups IA and IB according to CP and MP genes, but in subgroup IA according to the 2b gene. These data suggest that reassortment may have been a major event in the evolution of CMV in Iran, and that the Iranian isolates are derived from a common recent ancestor that had passed through a bottleneck event.
Koolivand, Davoud,Bashir, Nemat Sokhandan,Behjatnia, Seyed Aliakbar,Joozani, Raziallah Jafari The Korean Society of Plant Pathology 2016 Plant Pathology Journal Vol.32 No.5
The genomic region of Grapevine fanleaf virus (GFLV) encoding the movement protein (MP) was cloned into pET21a and transformed into Escherichia coli strain BL21 (DE3) to express the protein. Induction was made with a wide range of isopropyl-${\beta}$-D-thiogalactopyranoside (IPTG) concentrations (1, 1.5, and 2 mM) each for duration of 4, 6, or 16 h. However, the highest expression level was achieved with 1 mM IPTG for 4 h. Identity of the expressed protein was confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting. The expressed 41 kDa protein was purified under denaturing condition by affinity chromatography, reconfirmed by Western blotting and plate-trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA) before being used as a recombinant antigen to raise polyclonal antibodies in rabbits. Purified anti-GFLV MP immunoglobulines (IgGs) and conjugated IgGs detected the expressed MP and GFLV virions in infected grapevines when used in PTA-ELISA, double antibody sandwich-ELISA, and Western blotting. This is the first report on the production of anti-GFLV MP polyclonal antibodies and application for the virus detection.
Sevil Nematollahi,Nemat Sokhandan-Bashir,Farshad Rakhshandehroo,Hamid Reza Zamanizadeh 한국식물병리학회 2012 Plant Pathology Journal Vol.28 No.4
Molecular characterization of Cucumber mosaic virus (CMV) was done by using samples from tomato and cucurbitaceous plants collected from different locations in the northwest region of Iran. After screening by enzyme-linked immunosorbent assay, 91 CMV-infected samples were identified. Biological properties of eight representative isolates were compared with each other revealing two distinct phenotypes on squash and tomato plants. Phylogenetic analyses based on nucleotide sequences of the coat protein (CP), movement protein (MP) and 2b of the new isolates, together with that of previously reported isolates, led to the placement of the Iranian isolates in subgroups IA and IB according to CP and MP genes, but in subgroup IA according to the 2b gene. These data suggest that reassortment may have been a major event in the evolution of CMV in Iran, and that the Iranian isolates are derived from a common recent ancestor that had passed through a bottleneck event.
Davoud Koolivand,Nemat Sokhandan Bashir,Seyed Aliakbar Behjatnia,Raziallah Jafari Joozani 한국식물병리학회 2016 Plant Pathology Journal Vol.32 No.5
The genomic region of Grapevine fanleaf virus (GFLV)encoding the movement protein (MP) was clonedinto pET21a and transformed into Escherichia colistrain BL21 (DE3) to express the protein. Inductionwas made with a wide range of isopropyl-β-D-thiogalactopyranoside(IPTG) concentrations (1, 1.5, and2 mM) each for duration of 4, 6, or 16 h. However,the highest expression level was achieved with 1 mMIPTG for 4 h. Identity of the expressed protein wasconfirmed by sodium dodecyl sulphate polyacrylamidegel electrophoresis (SDS-PAGE) followed by Westernblotting. The expressed 41 kDa protein was purifiedunder denaturing condition by affinity chromatography,reconfirmed by Western blotting and platetrappedantigen enzyme-linked immunosorbent assay(PTA-ELISA) before being used as a recombinant antigento raise polyclonal antibodies in rabbits. Purifiedanti-GFLV MP immunoglobulines (IgGs) and conjugatedIgGs detected the expressed MP and GFLV virionsin infected grapevines when used in PTA-ELISA,double antibody sandwich-ELISA, and Western blotting. This is the first report on the production of anti-GFLV MP polyclonalantibodies and application forthe virus detection.