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

ABSTRACT: 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.

SUBMITTER: Lin B 

PROVIDER: S-EPMC4127255 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature