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Production of polyclonal phages harbouring antibody fragment genes against Xanthomonas citri subsp. citri using phage display technology

Document Type : Plant Pathology

Authors

1 Plant protection,agriculture,guilan university

2 Department of Plant Viruses, Iranian Research Institute of Plant Protection, Agricultural research education and extension organization of Iran. Tehran

3 Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran

4 Department of Plant Protection, College of Agricultural Sciences, Guilan University, Rasht

5 Departement of Plant Sciences & Biotechnology, Faculty of Life Sciences & Biotechnology, Shahid Beheshti University G.C, Tehran

Abstract

Citrus bacterial canker, caused by Xanthomonas citri subsp. citri (Xcc),is amongst the important diseases of lime orchards in southern parts of Iran. Phage display has been used to produce specific antibodies for detection of pathogen-infected plants as well as development of resistant varieties. An effector, namely pthA and a pilus protein, HrpE, the major critical components of type III secretion (T3S) system with roles in pathogenesis, were chosen as antigens. Recombinant forms of the proteins (pthA and HrpE) were expressed in a bacterial host and purified via affinity chromatography. Tomlinson phage display libraries including single chain variable fragments were used for isolation of the specific antibodies. Biopanning, 3 rounds against pthA and HrpE proteins, allowed enriching antigen-specific phages. The specificity of phages was tested using ELISA. Moreover, the phages were able to detect the plants infected with citrus bacterial canker.

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References
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Applied Entomology and Phytopathology
Volume 85, No. 2 - Serial Number 105
March 2018
Pages 265-276
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