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Creative Biolabs can provide Mempro™ cell-free membrane protein production services using baculovirus. Using our baculovirus expression system we can initiate the appropriate protocol for your custom project and then obtain the highest production of your target membrane protein.
Baculoviruses are known to infect invertebrates, with over 600 host species having been described. Immature (larval) forms of moth species are the most common hosts. Several GPCRs such as the ETB receptor has been expressed in E. coli at low yields due to its prokaryotic nature, possibly altering GPCR properties. The insect cell based on baculoviral has emerged as a popular membrane protein production system for rapid and high yield of several GPCRs.
In Mempro™ baculorivus-based cell-free protein production system, the protein production produces from infection of insect cells by recombinant viruses encoding the target gene of membrane protein. Unlike E. coli cell-free expression system, the Mempro™ baculovirus-based cell-free protein production system is an eukaryotic expression system. Additionally, this system uses a helper-independent virus that can be propagated to high concentration in insect cells adapted for growth in suspension cultures, making it possible to obtain large amounts of recombinant membrane proteins (Figure 1).
Figure 1. Recombinant protein expression in insect cells using baculovirus expression system.
Currently, the most widely employed baculovirus expression system is Autographa californica nucleopolyhedrovirus (AcNPV). Baculoviruses have biphasic life cycle, thus, two different types of virus particles are generated in the infection cycle. The first phase (10-24 h postinfection) involves the formation extracellular budded virus particles (BV), and the second phase (24-72 h postinfection) can generate large viral occlusion bodies (occluded virus, OV) from polyhedrin. The fact that polyhedrin is not essential for productive infection in vitro make baculoviruses desirable vectors for foreign gene expression. For recombinant membrane protein production, the polyhedrin gene of the viral DNA is substituted by the target gene of membrane protein, which is then expressed in high amounts. Then the functional baculovirus is propagated in insect cells to generate high concentration virus stocks that can be used for membrane protein production.
Therer are several advantages of Mempro™ cell-free protein production in baculovirus, including:
For clients who prefer a eukaryotic expression system, are exploring lower-cost alternatives to mammalian expression system but do not want to compromise on the overall quality of their recombinant membrane proteins, we strongly recommend considering our Mempro™ baculovirus-based cell-free membrane protein production service.
T. Lahtinen, et al. (2008). Hydrophobin (HFBI): A potential fusion partner for one-step purification of recombinant proteins from insect cells. Protein Expr. Purif., 59(1): 18-24.
Heterologous expression of the ETB receptor using the baculovirus/insect cells system. (http://www.diss.fu-berlin.de/diss/servlets/MCRFileNodeServlet/FUDISS_derivate_000000002653/2_kapitel2.pdf;jsessionid=16755B21E7EBA6A105C433AB04BD813B?hosts=)
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