The Expression Of Foot-and-Mouth Disease Virus Structural Protein VP1 In Transgenic Carrots

Foot-and-mouth disease is a kind of infectious disease of artiodactyls,which is acute,pyretic and high-tangent. There has been no effective method by far to cure this disease. Oralvaccines produced by transgenic plants are believed promising in the prevention of thisdisease. The carrot is apt to tissue culture and transformation. At the same time, it growswidely and can eat directly .Furthermore,it is perfect to men specially to babies. So it is aperfect "bio-reactor"in producing oral vaccines. In this work,Carrot was chose as thetransformation adaptor. FMDV structural protein VP1 gene was investigated,which wasexpressed in transgenic carrot. All of above provided a new approach of investigating thenew-style vaccine to defend and cure the foot-and-mouth disease.In the experiment,we firstly established a effective regeneration system of carrot byusing hypocotyls and cotyledons as explants from 3 carrot (Daucus carota L.var.sativa)breeding lines.On the basic of it,hypocotyls and calli of carrot were infected with Agrobacteriumtumefaciens LBA4404 harboring pBI121. By using X-Gluc confirmed that the transient andstable expression of GUS gene was obtained.Secondly,plant expressed vector pBFMDV which containing FMDV structural proteinVP1 gene under the control of 35S double promoter of cauliflower mosaic virus(CaMV) wasconstructed. By infected with Agrobacterium tumefaciens LBA4404,carrot cells weretransferred into FMDV VP1 gene. PCR and PCR-Southern analyses of carrot DNA confirmedthat the FMDV VP1 gene was introduced into the plant genome successfully. The result ofSDS-PAGE and Western-blotting showed that the gene of interest was expressed in some ofthe transgenic plants. The binding activity of the expressed proteins to anti-FMDV antibodieswas confirmed by using Dot-ELISA and ELISA. These results demonstrated that FMDVstructural protein VP1 had been expressed in transgenic carrots successfully,and theexpressed protein had activity .This experiment was the first report of producing FMDV vaccine in transgenic carrots.All the results will provide a new idea and a developmental foreground in producing oralvaccine of foot-and-mouth disease in transgenic plants.