Related Experiment Video
Updated: May 1, 2026
A Method For Production of Recombinant mCD1d Protein in Insect Cells.
Published on: December 10, 2007
Construction of recombinant bacmid containing m2e-ctxb and producing the fusion protein in insect cell lines
Nima Mirzaei1, Talat Mokhtari Azad2, Rakhshandeh Nategh2
1Department of Biology, Science and Research branch, Islamic Azad University, Tehran, IR Iran.
Background:
Sequence variations in glycoproteins of influenza virus surface impel us to design new candidate vaccines yearly. Ectodomain of influenza M2 protein is a surface and highly conserved protein. M2e in influenza vaccines may eliminate the need for changing vaccine formulation every year.
Objectives:
In this study, a recombinant baculovirus containing M2e and cholera toxin subunit B fusion gene was generated with transposition process to express in large amounts in insect cell lines.
Materials And Methods:
M2e-ctxB fusion gene was created and cloned into pFastBac HT. The recombinant vector was transformed into DH10Bac cells to introduce the fusion gene into the bacmid DNA via a site-specific transposition process. The recombinant bacmid was then extracted from white colonies and further analyzed using PCR, DNA sequence analyzing, and indirect immunofluorescence assay.
Results:
PCR and DNA sequence analyzing results showed that the fusion gene was constructed as a single open reading frame and was successfully inserted into bacmid DNA. Moreover, indirect immunofluorescence results showed that the fusion gene was successfully expressed.
Conclusions:
Baculovirus expression vector system is valuable to produce M2e based influenza vaccines due to its simple utilization and ease of target gene manipulation. The expressed protein in such systems can improve the evaluating process of new vaccination strategies.
