Dear Community, My apology for the off-topic question!
I am trying to express a 65 kDa human protease in insect cells using baculovirus expression system (Invitrogen). I used HBM secretion signal sequence to expression my protein of interest. Virus generation and titer amplification was done in Sf9 cells, and the protein was expressed in Hi5 cells. The western blot shows the expression of protein in the cells, but not in the supernatant, that means the protein is not secreted (I fear). Though it should be secreted in the medium as the HBM secretion signal sequence (Invitrogen) was there at the N-terminus. Also, my protein is a secreted protein, but I replaced the native signal peptide with the HBM signal sequence fearing that the mammalian signal peptide might not work in insect cell (though I am not sure)? If anybody has experienced the same, please share your experience and suggest me the further steps. Is it possible that the native signal is necessary for the secretion, even in the insect cell expression? Also what other signal sequences could be used instead of HBM? Thanks in advance for your suggestions/comments! Best, Dipankar -- *Dipankar Manna, Ph.D* Postdoctoral Researcher Department of Molecular Medicine Institute of Basic Medical Sciences University of Oslo, Domus Medica Oslo, Norway Mob : +47 451 66 517 E-mail: [email protected] <[email protected]> [email protected] http://www.med.uio.no/imb/english/people/aca/dipankam/
