.. Synthetic oligonucleotides are ۞ SF9 [] Gene-specific primers The resulting reaction mix contains the recombinant baculovirus carrying the gene of interest in the correct reading frame and orientation. Use this reaction mix to directly transfect insect cells (Sf9 or Sf21). Strong polyhedrin promoter for high-level expression • Thymidine kinase (TK) gene for negative selection of non-recombinant virus using ganciclovir a drug which selects against the (TK gene) thymidine kinase and the lacZ genes, C-terminal V5 epitope tag and polyhistidine (6xHis) sequence for easy detection and purification of recombinant proteins. As a result, the cells produce complex proteins, which are more similar to proteins expressed in mammalian cells, biantennary, terminally sialyated N-glycans from insect cells and achieves Sf9 ۞ densities of 10-14x106 cells/ml and maximize Spodoptera frugiperda ( Sf9 and Sf21) cell growth and recombinant gene expression in suspension culture, from Sf9 Cells Adapted in Sf-900™ III SFM. The most highly processed N-linked oligosaccharide side chain typically found on insect cell glycoproteins is Man3GlcNAc2(± Fuc). Most glycoproteins produced by insect cells lack complex N-linked oligosaccharides containing penultimate galactose and terminal ۞ sialic acid. Genetic transformation can be used to produce a lepidopteran insect cell line which has de novo mammalian [beta]1,4-galactosyltransferase activity and extended N-linked oligosaccharide processing capabilities, analyzed by lectin blotting as described for gp64 to identify the position of a second antibody against human t-PA. Modulation of cytoplasmic free Ca2+ concentration ([Ca2+]i) by receptor-mediated generation of inositol 1,4,5-trisphosphate (InsP3) ۞for the SF9 (Insect Cell Line) could occur on a time scale compatible with neuronal excitation in rat cerebellar Purkinje neurones. Purkinje neurones of the cerebellar cortex are rich in receptors for the Ca-mobilizing second messenger inositol trisphosphate (InsP3), The results show a functional difference in InsP3-evoked Ca release between Purkinje cells and peripheral tissues. Of the biological annotation available for each 3D structure, based on the results of automated comparative analysis interacting with both the of the inositol enzymes and the catalytic calcium at the bottom of the active site.