| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | For translation efficiency in a eukaryotic expression system, the Kozak sequence (CCACCAUGG) was inserted and for purification of the protein, a polyhistidine-tag was added at the 5' end of the gene sequence. The gene sequences was completely synthesized and cloned into Puc19 vector by GenScript, USA, Inc. The synthetized Beclin1 gene was digested from Puc19 vector using suitable restriction enzymes and was sub-cloned into multiple cloning site 1 of the eukaryotic expression vector pVITRO2-neo-mcs (InvivoGen, France). | Get A Quote |
Introduction: Autophagy is a complicated process which is involved in many biological events such as antigen presentation by immune cells. Beclin1, as a key component of autophagic machinery, plays the main role in the induction and initiation of this process. In the present study, we hypothesized that overexpression of Beclin1 could be useful in autophagy induction as an immunostimulatory strategy for improving the efficiency of DNA vaccines. Methods: Beclin1 gene was cloned into pVITRO2 eukaryotic expression vector and was confirmed by agarose gel electrophoresis. Beclin1 expression was evaluated by Real-Time RTPCR and Western blotting. The autophagy induction by pVITRO2 encoding Beclin1 in HEK293 cells was e... More
