Most membrane proteins are not sufficiently abundant in nature to purify and crystallize, and gene expression systems have been unable to reliably express membrane proteins in sufficient quantities. The baculovirus expression vector system (BEVS) is one of the most widely used gene expression systems for eukaryotic proteins and is optimal for glycosylated protein expression in a cost-effective manner. BEVS has also been used to express some membrane proteins. Despite some success, however, many membrane proteins cannot be expressed at high enough levels for structural studies. One possible means for improving membrane expression using BEVS is to develop new cell lines with greater intrinsic ability for membrane protein synthesis.
Michigan State University’s invention is a new cell line derived from Trichoplusia ni embryos for use in BEVS known as TnT4. TnT4 has favorable growth characteristics for use in BEVS and expresses a human neurotensin receptor 1 (NTSR1), a model G-protein coupled receptor with the ability to express membrane proteins at higher levels. TnT4 cells have been shown to express the membrane-GFP-fusion protein at approximately twice the level of Sf21 cells, as evaluated by GFP intensity. They are susceptible to infection by Autographa californica nucleopolyhedrovirus (AcMNPV) and support its complete lifecycle, producing both BV and polyhedra. The cell line develops rapidly and can be grown in suspension culture, providing favorable growth characteristics for both research and drug development.
Benefits
* Free of Latent Nodavirus
* Improved membrane protein expression: Expression of the membrane-GFP-protein at levels twice that of the Sf21 cell line has been demonstrated.
* Compatible with existing BEVS technologies: The cell line can be used with little or no changes in current methodologies.
* Favorable growth: Grows in suspension culture with short doubling time.
Applications
* Drug and vaccine development and production
* Diagnostic test development
* Insecticide development
* Three-dimensional structural protein research
IP Protection Status
U.S. Patent Issued: 8,383,402
Licensing
This technology is licensed on a non-exclusive basis
