Purification and immuno-gold labeling of lily mottle virus from lily leaves

Yubao Zhang; Yajun Wang; Zhongkui Xie; Ruoyu Wang; Zhihong Guo; Wopke van der Werf; Le Wang

doi:10.1016/j.jviromet.2016.02.001

Purification and immuno-gold labeling of lily mottle virus from lily leaves

Yubao Zhang, Yajun Wang, Zhongkui Xie^*, Ruoyu Wang, Zhihong Guo, Wopke van der Werf, Le Wang

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

5 Citations (Scopus)

Abstract

Lily mottle virus (LMoV) is prevalent in Lilium species worldwide causing dwarfing, flower breaking, and reduced bulb yield. In this paper, an easy to use and efficient procedure is described for purification of LMoV from lily leaves. The resulting sample is characterized by a 260/280 nm absorbance ratio of 1.20 at a concentration of 1.27 mg/ml. The procedure results in high protein purity and particle integrity as shown by UV-spectrophotometry, polyacrylamide gel electrophoresis (PAGE), Western blotting, reverse transcriptase (RT)-PCR and transmission electron microscopy (TEM) in combination with immuno-gold labeling. This is the first time that an immuno-gold labeling (IGL) assay was performed to identify a virus of lily. Purified products can be used as a source of antigen in the preparation of antibodies against LMoV and may assist in the development of a diagnostic test for LMoV and in epidemiological surveys.

Original language	English
Pages (from-to)	33-38
Journal	Journal of Virological Methods
Volume	232
DOIs	https://doi.org/10.1016/j.jviromet.2016.02.001
Publication status	Published - 2016

Keywords

Immuno-gold labeling assay
Lily mottle virus
Purification
Transmission electron microscope

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@article{f3574a6fa07741f196e55f26aab0c63b,

title = "Purification and immuno-gold labeling of lily mottle virus from lily leaves",

abstract = "Lily mottle virus (LMoV) is prevalent in Lilium species worldwide causing dwarfing, flower breaking, and reduced bulb yield. In this paper, an easy to use and efficient procedure is described for purification of LMoV from lily leaves. The resulting sample is characterized by a 260/280 nm absorbance ratio of 1.20 at a concentration of 1.27 mg/ml. The procedure results in high protein purity and particle integrity as shown by UV-spectrophotometry, polyacrylamide gel electrophoresis (PAGE), Western blotting, reverse transcriptase (RT)-PCR and transmission electron microscopy (TEM) in combination with immuno-gold labeling. This is the first time that an immuno-gold labeling (IGL) assay was performed to identify a virus of lily. Purified products can be used as a source of antigen in the preparation of antibodies against LMoV and may assist in the development of a diagnostic test for LMoV and in epidemiological surveys.",

keywords = "Immuno-gold labeling assay, Lily mottle virus, Purification, Transmission electron microscope",

author = "Yubao Zhang and Yajun Wang and Zhongkui Xie and Ruoyu Wang and Zhihong Guo and {van der Werf}, Wopke and Le Wang",

year = "2016",

doi = "10.1016/j.jviromet.2016.02.001",

language = "English",

volume = "232",

pages = "33--38",

journal = "Journal of Virological Methods",

issn = "0166-0934",

publisher = "Elsevier",

}

TY - JOUR

T1 - Purification and immuno-gold labeling of lily mottle virus from lily leaves

AU - Zhang, Yubao

AU - Wang, Yajun

AU - Xie, Zhongkui

AU - Wang, Ruoyu

AU - Guo, Zhihong

AU - van der Werf, Wopke

AU - Wang, Le

PY - 2016

Y1 - 2016

N2 - Lily mottle virus (LMoV) is prevalent in Lilium species worldwide causing dwarfing, flower breaking, and reduced bulb yield. In this paper, an easy to use and efficient procedure is described for purification of LMoV from lily leaves. The resulting sample is characterized by a 260/280 nm absorbance ratio of 1.20 at a concentration of 1.27 mg/ml. The procedure results in high protein purity and particle integrity as shown by UV-spectrophotometry, polyacrylamide gel electrophoresis (PAGE), Western blotting, reverse transcriptase (RT)-PCR and transmission electron microscopy (TEM) in combination with immuno-gold labeling. This is the first time that an immuno-gold labeling (IGL) assay was performed to identify a virus of lily. Purified products can be used as a source of antigen in the preparation of antibodies against LMoV and may assist in the development of a diagnostic test for LMoV and in epidemiological surveys.

AB - Lily mottle virus (LMoV) is prevalent in Lilium species worldwide causing dwarfing, flower breaking, and reduced bulb yield. In this paper, an easy to use and efficient procedure is described for purification of LMoV from lily leaves. The resulting sample is characterized by a 260/280 nm absorbance ratio of 1.20 at a concentration of 1.27 mg/ml. The procedure results in high protein purity and particle integrity as shown by UV-spectrophotometry, polyacrylamide gel electrophoresis (PAGE), Western blotting, reverse transcriptase (RT)-PCR and transmission electron microscopy (TEM) in combination with immuno-gold labeling. This is the first time that an immuno-gold labeling (IGL) assay was performed to identify a virus of lily. Purified products can be used as a source of antigen in the preparation of antibodies against LMoV and may assist in the development of a diagnostic test for LMoV and in epidemiological surveys.

KW - Immuno-gold labeling assay

KW - Lily mottle virus

KW - Purification

KW - Transmission electron microscope

U2 - 10.1016/j.jviromet.2016.02.001

DO - 10.1016/j.jviromet.2016.02.001

M3 - Article

AN - SCOPUS:84960918878

VL - 232

SP - 33

EP - 38

JO - Journal of Virological Methods

JF - Journal of Virological Methods

SN - 0166-0934

ER -