Applying of recombinant protein technology fordevelopingof specific antibodyagainst Citrus tristeza virus

Background and Objective: Citrus tristeza virus (CTV) is one of the most destructive viruses that causea lot of damage to agricultural products all over the world every year. Therefore, it is important to detectthis virus in plant samples and ensure the delivery of healthy inputs to farmers. One of the usefuldiagnostic methods is the ELISA technique, which requires the use of an antibody against the coat proteinof the virus. Therefore, the aim of this research is to produce polyclonal antibodies for the preparation ofvarious disease diagnostic kits.Methods: In this research, first the coat protein gene of the virus was isolated from the infected samplesand then the corresponding gene was cloned in the pET ۲۸ a(+) expression vector. In the next step, theexpression vector containing the desired gene was introduced into the BL۲۱ strain E. coli bacterium toexpress the coat protein of the Tristeza virus in this bacterium. After the production of the mentionedprotein, which has a histidine tag, purification was done using chromatography. Then the purified proteinwas injected into the rabbit to produce antibodies against it in the rabbit's blood. In the last stage, Elisa,Western Blot, and DBIA methods were used to check the sensitivity and specificity of the producedantibody.Results: The obtained results show the good quality of the produced antibody, which can be used in fieldand laboratory conditions to identify plants infected with citrus tristeza virus.Conclusion: According to the obtained results, it seems that the internal production of this diagnostic kitcan significantly help to reduce the diagnosis costs and prevent the spread of the mentioned disease in thecitrus gardens of the country.

Keywords:

Citrus tristeza virus , DBIA , Polyclonal antibody , Western blot