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   Developing of specific antibody against chickpea chlorotic dwarf virus (CpCDV) through recombinant coat protein  
   
نویسنده safarnejad mohammad reza ,bananej kaveh ,sokhansanj yalda
منبع journal of crop protection - 2019 - دوره : 8 - شماره : 2 - صفحه:179 -190
چکیده    The legume crops such as chickpea and lentils are mainly cultivated in semi-arid tropical lands. chickpea chlorotic dwarf virus (cpcdv) causes major losses to legumes throughout the world. producing of specific antibody against this virus is crucial for surveys of disease in the fields and assessment of vial resistance in plant cultivars. present article describes developing of specific antibody against the cpcdv virus by applying recombinant protein. in this study, coat protein of cpcdv was selected as a target for detection and preparation of polyclonal antibody. to achieve this aim cp gene encoding coat protein of cpcdv was initially pcr-amplified and inserted into bacterial expression vector. expression of recombinant protein was performed in bl21 strain of escherichia coli. purification was carried out under native conditions and the accuracy of recombinant protein production was confirmed by electrophoresis. the purified recombinant coat protein of cpcdv was used forimmunization of rabbit. purification of immunoglobulin molecules wasperformed by affinity chromatography using protein a column followed byconjugating of igg to alkaline phosphatase enzyme. the capability of purifiedantibodies and conjugates for efficient detection of infected plants wasassessed by double antibody sandwich enzyme-linked immunosorbent assay (das-elisa), western blotting and dot immunosorbent assay (diba). these results proved that prepared igg and conjugate are able to distinguish with high efficiency cpcdv infected plants. to the best of our knowledge, this is the first report for production of anti-cpcdv antibodies raised through recombinant protein technology.
کلیدواژه antibody ,chickpea ,CpCDV ,ELISA ,recombinant protein
آدرس agricultural research, education and extension organization (areeo), iranian research institute of plant protections, Iran, agricultural research, education and extension organization (areeo), iranian research institute of plant protections, Iran, islamic azad university, tehran science and research branch, faculty of agriculture and natural resources, department of plant pathology, Iran
 
     
   
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