Expression And Variance Analysis Of FMDV Combining Gene In Bombyx Mori Baculovirus Expression Vector System

Foot-and-mouth disease (FMD) is an extremely contagious animal disease caused by foot-and-mouth disease virus (FMDV)which mainly infects cloven-hoofed animals such as swine,cattle and sheep. FMD seriously impair healthy development of animal husbandry and result in great economic loss to the national economy. Therefore,all the countries pay much attention to its study and prevention around the world. Now, the traditional vaccines still remains as the most preferred route of administration in prevention and control of FMD.However,it is difficult to avoid the expensive production and the potential risk factors such as the escaping virus in the process of preparing vaccines. Therefore, a safe and highly efficient FMD molecule vaccine is in great need. In this study,the type 0 FMDV combination of genes was expressed in Bombyx mori baculovirus expression system and the expressions of combining gene of AsiaI FMDV in different species of silkworm were analyzed, which will provide references for the production of FMD molecule vaccines with the use of Bombyx mori Bioreactor.(1) In this experiment,we selected FMDV capsid protein procusor P1 and protease 2A,3C of OMIII strain,which were inserted into Bombyx mori baculovirus transfer vector pVL1393 and constructed the recombinant vector pVL-P12A3C, with which co-transfected linearted DNA of Bm-BacPAK6 virus into BmN cells. The recombinant virus Bm-P12A3C was gained. Indirect immunofluorescence test determined that recombinant virus (Bm-P12A3C) could validly express the target gene in Bm-N cell. At the same time, the fifth instars were infected with the recombinant virus. The target protein in haemolymphe was expressed according to double-antibody sandwich ELISA result. Screening and purification of recombinant virus,with which the fifth instars were infected again, detecting them by the same method. The result indicated that the high expression of FMDV antigen has been obtained in Bombyx mori baculovirus expression system.(2) Selecting 20 species of silkworm pupa and injecting the recombinant virus which had received the stable expression of P1-2A 3C protease coding regions of FMDV Asia I.The target protein in silkworm pupa was used to determine by indirect sandwich ELISA and the variance analysis was made. The result indicated that there was certain difference in these varieties, in which the No.of 9,10,12,14,15,17 were relatively higher expression.