9058009712

enveiope pioiem wiis piuuuceu nom L>acuiovirus

Exprcssion Vcctor System known as one of the most powerful and versatile eukaryote expression vector system available. It has been used to express many prołeins from different sources. High level of protein production and posttranslational modification of recombinant protein can be obtained using insect celt as host for baculovirus replication. Insect eelts were used as hosts for production of recombinant emelope protein when infected with recombinant emelope bacutovirus that was engineered by insertion of em elope gene under control of the strong polyhedrin promoter. The recombinant E protein can be used in the diagnostic system as an antigen to detect dengue viral infection. Recombinant protein production is affected by several factors such as celt density, time of infection (TOI) and availabi 1 ity of nutrients including multiplicity of infection (MOI) and dissoWed oxygen.

exponentiai pnase (l and z*iu ceus/mi) witn recombinant F. baculovirus at various MOI of

0. 1, 1,5 and 10. One ml of infected celi was collected cłaily for celi count while one ml was centrifuged at 10,000 rpm for 10 min, celi pellel was collected and resuspended in celi lysis buffer. Recombinant dengue v i ra 1 emelope protein from celi lysate was detected by Western biot analysis. The intensity of spec i He protein bands recognized by monoclonal antibody (3115) was quantified by image analyzer against purified recombinant dengue viral em elope protein from E.coli.

Rcsults and Discussion:

1.    Growth medium: Ten different media were tested by culturing SfO cclls in 250 ml shake fiask. The maximum viable celi density and the pricc of medium were compared in Table 1.