preparation of antibody against immunodominant membrane protein (imp) of candidatus phytoplasma aurantifolia
نویسندگان
چکیده
background: the witches’ broom disease of lime caused by candidatus phytoplasma aurantifolia, is the most devastating disease of acidian lime in the southern parts of iran. objectives: at present, no effient method has been developed for controlling the disease, therefore quarantine approaches such as early detection and subsequent eradication of infected trees is very important. toward this aim, developing a reliable and sensitive detection method would be the fist step to prevent transportation of infected plant materials to other places. background: the witches’ broom disease of lime caused by candidatus phytoplasma aurantifolia, is the most devastating disease of acidian lime in the southern parts of iran. objectives: at present, no efficient method has been developed for controlling the disease, therefore quarantine approaches such as early detection and subsequent eradication of infected trees is very important. toward this aim, developing a reliable and sensitive detection method would be the first step to prevent transportation of infected plant materials to other places. materials and methods: in this study, immunodominant membrane protein (imp) of the pathogen was selected as a target for detection and preparation of polyclonal antibody. the imp is the major protein present on the surface of phytoplasma cells. for this purpose, the dna region encoding imp gene was isolated and cloned into pet28a bacterial expression vector. the recombinant protein was expressed in a large scale in escherichia coli. purification was performed under native conditions and the purity and integrity of produced recombinant protein were confirmed by western immuno blot analysis using anti his-tag and anti-imp polyclonal antibodies. the purified recombinant imp was used for immunization of rabbit. purification of immunoglobulin was performed by affinity chromatography using protein a column. the purified immunoglobulin was conjugated with the alkaline phosphatase enzyme. results: the purified antibodies and conjugates were applied for efficient detection of infected plants in double antibody sandwich enzyme-linked immunosorbent assay (das-elisa) and dot immunosorbent assay (diba). conclusions: these antibodies were proven to be very powerful tools to detect the candidatus phytoplasma aurantifolia in plants.
منابع مشابه
Preparation of Antibody Against Immunodominant Membrane Protein (IMP) of Candidatus Phytoplasma aurantifolia
Background: The witches’ broom disease of lime caused by <span style="font-variant: normal; font-style: normal; font-family: Capitolium...
متن کاملGeneration and expression in plants of a single-chain variable fragment antibody against the immunodominant membrane protein of Candidatus phytoplasma aurantifolia.
Witches' broom of lime is a disease caused by Candidatus Phytoplasma aurantifolia, which represents the most significant global threat to the production of lime trees (Citrus aurantifolia). Conventional disease management strategies have shown little success, and new approaches based on genetic engineering need to be considered. The expression of recombinant antibodies and fragments thereof in ...
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Witches' broom disease of acid lime greatly affects the production of Mexican lime in Iran. It is caused by a phytoplasma (Candidatus Phytoplasma aurantifolia). However, the molecular mechanisms that underlie phytoplasma pathogenicity and the mode of interactions with host plants are largely unknown. Here, high-throughput transcriptome sequencing was conducted to explore gene expression signatu...
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متن کاملارزیابی واکنش تعدادی از هیبریدهای مرکبات نسبت به 'Candidatus Phytoplasma aurantifolia'
مناطق جنوبی ایران شامل استانهای هرمزگان، سیستان و بلوچستان، بوشهر، فارس و کرمان سابقهای طولانی در تولید لیموترش (مکزیکن لایم) [Citrus aurantifolia (Chrism.) Swing.] دارند. در دو دهه اخیر بیماری جاروک لیموترش که عامل آن 'Candidatus Phytoplasma aurantifolia' میباشد هزاران اصله از درختان لیموترش را در جنوب ایران نابود کرده است. یکی از راههای مدیریت بیماری جاروک لیموترش به کار...
متن کاملIn vitro interactions between immunodominant membrane protein of lime witches’ broom phytoplasma and leafhopper vector proteins
Lime witches’ broom phytoplasma (16SrII, LWB) causes major losses to lime cultivation in Iran. The role of its immunodominant membrane protein (IMP) in the specific interaction with vector proteins was addressed. The carboxy terminal moiety of LWB IMP, coding the extracellular part of the protein, was expressed (LWB f∆IMP) as a tool to study IMP interactions with insect proteins. In preliminary...
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عنوان ژورنال:
iranian journal of biotechnologyناشر: national institute of genetic engineering and biotechnology
ISSN 1728-3043
دوره 11
شماره 1 2013
کلمات کلیدی
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