The Study On High-level Expression Of _HuIFN_α-_HuTHY_α1,_HuIFN_β,HuIFN_β Mutation In Silkworm Expression System And Bioactivities Of Products

1 The baculovirus shuttle vector , pBacPAK-IFN-THY was constructed which contains the gene of IFN- a 2b and THY- a 1. Constructed vector Was coinfected with linear Bm-BacPAK6 DNA into BmN Cells. The recombinant virus was screened and plaque-purified. Fusion gene was successfully expressed in BmN cells, silkworm larvae and pupae. The results showed that protein was successfully prepared and its bioactivities were testified by WISH-VSV system and ERFC. Results indicated the expressed fusion protein have bioactivities of both IFN- a 2b and THY- a 1. Furthermore, ELISA assay showed that fusion protein IFN-THY expressed in silkworm pupae can be absorbed into the blood stream of mice administered by stomach perfusion.Two baculovirus shuttle vectors, pBacPAK-IFN B and pBacPAK-IFN B s17 was respectively constructed which contains IFN B and the mutant gene of IFN B S17. Constructed vector was coinfected with linear Bm-BacPAK6 DNA into BmN Cells. The recombinant virus was screened and plaque-purified. The BmN cells and silkworm larvae were infected with the recombinant virus. The results showed that proteins was successfully prepared and their bioactivities were testified by WISH-VSV system. Results indicated the expressed recombinant proteins have full bioactivity of HuIFN B .