Production of Polyclonal Antibody Against Movement Protein of Tomato Yellow Leaf Curl Virus and Assessment of Its Efficacy in Virus Detection

Document Type : Research Paper

Authors

1 Former M.Sc. Student, Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran

2 Associate Professor, Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran

Abstract

Tomato yellow leaf curl disease (TYLCD) is one of the most important diseases of tomato (Solanum lycopersicum L.) plants in tropical, subtropical regions and greenhouses in temperate regions of the world. To produce an antibody against the viral agent of the disease for use in a diagnostic kits, the TYLCV cloned viral movement protein gene was expressed in an expression vector (pET26). The resulting recombinant protein was then injected into a rabbit. The prepared antiserum was used in western blot and ELISA analyses in order to assess its efficiency in virus detection. A 13KD protein band related to viral movement protein was detected in infected plants by using Western blot analysis at 1:1500 antiserum dilutions, while in ELISA tests at 1:1000 dilutions of antiserum, denaturation of the protein could increase the differences between the absorbance values of the infected plants and non-infected plants. However, these differences were not statistically significant. The results showed that the prepared antiserum against Tomato yellow leaf curl virus- movement protein was suitable for western blot analyses, but to be used in ELISA test the antibodies need to be extraction and purification of antibodies.

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References

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Volume 6, Issue 2
May 2016
Pages 33-37

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