High variability in the N terminus of coat protein among potyviruses and its advantage in producing a specific antibody
- 71 Downloads
Cross reaction often occurs in serological detection for potyviruses that can lead to false positive results. Sequence alignment revealed that the N-terminal sequences of coat proteins (CP) are highly variable among potyviruses. Based on this finding, the preparation of a specific antibody against potyviruses is described here increasing the efficiency of detection and identification. A case study of Turnip mosaic virus (TuMV): a specific fragment “CP-50” (1–50 aa of CP) was obtained by prokaryotic expression and used for producing the polyclonal antibody. Test of specificity indicated that the prepared antibody “Anti-CP-50” strongly reacted with TuMV and is appropriate as a specific anti-TuMV antibody.
KeywordsPotyviruses Turnip mosaic virus Antibody Prokaryotic expression
Turnip mosaic virus
Enzyme linked immunosorbent assay
Grand average of hydropathicity.
The authors thank Prof. Chang Chin-an (Graduate Institute of Biochemical Sciences and Technology, Chaoyang University of Technology, Taichung, 41349, Taiwan) for providing experimental materials and technical assistance. We thank Peter Sol Reinach (Wenzhou Medical University) for providing language editing service of this manuscript. This work is supported by the grant from Fujian Provincial Natural Science Foundation of China (2011D002) and Xiamen Municipal Natural Science Foundation (3502Z20127015 & 3502Z20162014).
YLM and QXT conceived the experiments, and critically revised the manuscript. GHZ collected the samples. DWP and GHZ performed the experiments. ZZZ analyzed the data. DWP wrote the manuscript.
Fujian Provincial Natural Science Foundation of China (2011D002); Xiamen Municipal Natural Science Foundation (3502Z20127015 & 3502Z20162014).
Compliance with ethical standards
This study was carried out following the guidelines and approval of the Laboratory Animal Ethics Committee of Wenzhou Medical University. The rabbits were anesthetized by 3% pentobarbital sodium (1 ml/kg) when collecting blood samples. After the experiments, rabbits were submitted to euthanasia using anesthetic overdose.
Conflict of interest
The authors declare that they have no competing interests.
- Cooper, H. M., & Paterson, Y. (1995). Assays for antibody production. Current Protocols in Immunology, 2(4), 1–2.4.9.Google Scholar
- Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, M. R., Appel, R. D., et al. (2005). Protein identification and analysis tools on the ExPASy server. In J. M. Walker (Ed.), The proteomics protocols handbook. Totowa: Humana Press.Google Scholar
- Hong, J., Li, D. B., & Zhou, X. P. (2001). Classification atlas of plant virus. Beijing: Science Press.Google Scholar
- ICTV. (2017). Virus Taxonomy: The Classification and Nomenclature of Viruses: The Online (10th) Report of the ICTV. http://talk.ictvonline.org/ictv-reports/ictv_online_report.
- Kantrong, S., Saunal, H., Briand, J. P., & Sako, N. (1995). A single amino acid substitution at N-terminal region of protein of Turnip mosaic virus alters antigenicity and aphid transmissibility. Virology, 140, 453–467.Google Scholar
- Naidu, R. A., & Hughes, J. d’A. (1997). Methods for the detection of plant virus diseases. International Institute of Tropical Agriculture, Ibadan, Nigeria, 45: 1–28.Google Scholar
- Shukla, D. D., Tribbick, G., Mason, T. J., Hewish, D. R., Geysen, H. M., & Ward, C. W. (1989b). Localization of virus-specific and group-specific epitopes of plant potyviruses by systematic immunochemical analysis of overlapping peptide fragments. Proceedings of the National Academy of Sciences of the United States of America, 86, 8192–8196.CrossRefPubMedPubMedCentralGoogle Scholar
- Zheng, G. H., Peng, D. W., Tong, Q. X., Zheng, Z. Z., & Ming, Y. L. (2017). Occurrence of Turnip mosaic virus in Phalaenopsis sp. in China. Journal of Plant Pathology, 99, 703–706.Google Scholar