Research On The Intracellular Immunization To Dengue Virus By The Mosquito, Aedes Albopictus

Dengue fever is one of the most pandemic arbovirus diseases all over the world, which is caused by Dengue virus and transmitted by mosquito vector, Aedes. The vector control is the most effective method to prevent and control DF. But the traditional vector control measures have a few of limitations. The transgenic technology provides a new strategy for the control of arthropod-borne diseases. In this research, we amplified and cloned C and prM gene of Dengue virus, constructed the expression vector pB[hspEGFP] . The prM gene was inserted into insect expression vector by three kinds of expression ways. The expression of prM-EGFP fusion protein in Aedes albopictus cell, C6/36 was tested by Western blot and fluorescent analysis. The C6/36 cells which were transfected by recombinant plasmid produced resistance to dengue virus invasion. The phenomena of intracellular immunization of C6/36 cell against dengue virus were replicated. The result testified that the transcription of prM gene would cause intracellular immunization, whether the prM protein was expressed or not. And intracellular immunization was induced by both sense and anti-sense transcription of prM gene. We inferred that the mechanism of intracellular immunization might be caused by RNA interference. We also constructed the transgenic plasmid pB[PUBnlsEGFP-prM] based on transposon piggy Bac element. The EGFP-prM gene was tested to integrate into C6/36 cell genome by PCR and Southern blot. This experiment demonstrated that transposon piggyBac element and promoter polyubiquitin could function at Aedes albopictus and were useful to further construct dengue-refractory transgenic Aedes albopictus.