PARATRANSGENESIS AND SIRNA AS A POTENT TOOLS TO FIGHT AGAINST MALARIA: RNC GENE MOLECULAR CHARACTRIZATION OF ASAIA SP. THE SYMBIONT AGENT OF ANOPHELES STEPHENSI

Background: Malaria is the most important protozoan infectious disease in human and World Health Organization have some objectives towards the elimination of Malaria till 2050. One of the main object is use the VIMT vaccines. Carboxypeptidase Enzymes have main role in digestion of proteins in the midgut of the Anopheles and interruption in the normal function of this enzymes may cause the significance effect on the plasmodium life cycle and anopheles fitness. It is expected that with the inhibition of simultaneous activity of both cb1&cb2 enzymes, the inhibition of their activity could be synergically increased. At this investigation we clone and express the arboxypeptidase B1 to produce large scale of the enzyme for future studies.Objective: cloning and expression of the Carboxypeptidase B1 of Anopheles Stephensi.Materials and methods: Asaia bacteria were isolated from the larval and adult’s midgut of An. stephensi. Firstly, based on the bioinformatic analysis, middle part of the gene was determined and then was amplified by PCR. In the second step, genome walking method was used to amplify the upstream and downstream of this middle part based on asymetric PCR.Results and Discussion: Full length sequence of the rnc gene was acheived by using the genome walking method. Charactrization of this gene is fundemental step to create the Rnase III mutant bacteria as a siutable strain for construction of dsRNA to target diffrent gene in the vector based on RNAi mechanism.Conclusion: As mentioned, RNase III mutant strains are more efficient in dsRNA production. Therefore, by performing this study, we identified the RNase III gene of Asaia bacteria which is the fundamental step for creating the Asaia RNase III mutant strain

Keywords:

Asaia , Malaria , paratransgenesis , Anopheles , Rnase III , RNAi , dsRNA production , Vector control