Split cell and non cell cracking baculovirus system performance comparison and study membrane proteins
Abstract
To investigate the cracking of non-cell baculovirus expression system advantages and feasibility of use in this study 5 '-BrdU
mutant recombinant viruses vABhcmEpL, and select the cell with the most obvious split of the non-recombinant virus strains C4, and polh (AcMNPV polyhedrin) promoter activity in most of the recombinant virus strain PN24, to explore the split of non-cell baculovirus
expression system and cell cracking baculovirus expression system.
vABhcmEpL Department of recombinant AcMNPV virus containing the CMV minimal promoter (promoter) start, provided by the hr enhancer (enhancer) activity (hCMVm) the EGFP gene, and starting from the polh promoter fluorescent enzyme (luciferase) gene. These two genes play a reporter gene in this role. The Arabidopsis (Arabidopsis thaliana) of the sodium ion channel proteins CHL1 of N-trans-membrane region (transmembrane domains) with the disk to be coral sea anemone
(Discosoma sp.) Red fluorescent protein DsRed bonding may be a fusion protein CHL1-DsRed. Then vABhcmEpL, C4, PN24 and other
recombinant virus again, from every strain of the virus in the build out of two recombinant viruses, respectively, with the hCMVm or polh promoter start of CHL1-DsRed protein. The infection of the Sf21 cells to microscopic observation and measurement of fluorescence
intensity with a fluorescence photometer, observing hCMVm effect with polh promoter and, using Western blot analysis of cell debris and supernatant. Test results showed membrane protein, the early start of production of child hCMVm late promoter polh than high;
rather than cell cracking baculovirus expression system than the traditional split of baculovirus cell expression system with high output.
