Enhancement of IgG Purification by FPLC for a Serological Study on the Turnip mosaic virus P1 Protein

The P1/HC-Pro of Potyvirus is the first discovered viral gene-silencing suppressor in which helper-component-proteinase (HC-Pro) function in concert with P1 to improve the suppression of post-transcriptional gene silencing (PTGS). However, the mechanism of P1/HC-Pro remains unclear. The P1 protein i...

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Published in植物病理學會刊 Vol. 22; no. 1; pp. 021 - 030
Main Authors 邱敏慈(Chiu,Ming-Tzu), 林長平(Lin,Chan-Pin), 林品均(Lin,Pin-Chun), 林詩舜(Lin,Shih-Shun)
Format Journal Article
LanguageChinese
Published 台灣 中華民國植物病理學會 01.03.2013
Subjects
fast protein liquid chromatography
FPLC
immunoglobulin G
P1
P1 蛋白
Turnip mosaic virus
免疫球蛋白 IgG
蕪菁嵌紋病毒
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ISSN1021-9544

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Summary:The P1/HC-Pro of Potyvirus is the first discovered viral gene-silencing suppressor in which helper-component-proteinase (HC-Pro) function in concert with P1 to improve the suppression of post-transcriptional gene silencing (PTGS). However, the mechanism of P1/HC-Pro remains unclear. The P1 protein is the most divergent protein regarding length and the amino acid sequence in potyviruses. An effective serological tool is required to examine the function of P1. To improve the titer of the antiserum to P1 of Turnip mosaic virus (TuMV), we developed a procedure using fast protein liquid chromatography (FPLC) to purify immunoglobulin G (IgG). Moreover, the purified IgGs were further concentrated using appropriate centrifugal filter device to enhance the sensitivity. The results indicated that the new procedure improved the efficiency of IgG purification in a few hours. The low P1 signal was difficult to be detected with unpurified antersum to P1 in the TuMV-infected N. benthamiana plants, whereas the purified IgG t
ISSN:1021-9544