Efficient expression of acetylcholine-binding protein from Aplysia californica in Bac-to-Bac system.

Lin B1, Meng H1, Bing H1, Zhangsun D1, Luo S1.
Source: Biomed Res Int.
Publication Date: (2014)
Issue: 691480: Epub 2014 Jul 20
Research Area:
Basic Research
Cells used in publication:
Species: spodoptera frugiperda (army fall worm)
Tissue Origin: ovarian
Culture Media:

Expression of Acetylcholine-binding Protein in Baculovirus System


The Bac-to-Bac baculovirus expression system can efficiently produce recombinant proteins, but the system may have to be optimized to achieve high-level expression for different candidate proteins. We reported here the efficient expression of acetylcholine-binding proteins from sea hares Aplysia californica (Ac-AChBP) and a convenient method to monitor protein expression level in this expression system. Three key factors affecting expression of Ac-AChBP were optimized for maximizing the yield, which included the cell density, volume of the infecting baculovirus inoculums, and the culturing time of postinfection. We have found it to reach a high yield of ~5 mg/L, which needs 55 h incubation after infection at the cell density of 2 × 10(6) cells/mL with an inoculum volume ratio of 1 : 100. The optimized expression system in this study was also applied for expressing another protein Ls-AChBP from Lymnaea stagnalis successfully. Therefore, this established method is helpful to produce high yields of AChBP proteins for X-ray crystallographic structural and functional studies.