TITLE:
Expression and localization of the spore wall protein SWP26 of Nosema bombycis in the silkworm BmN cell line
AUTHORS:
Feng Zhu, Zhongyuan Shen, Shengyan Xiao, Yajie Yue, Xuliang Fu, Xudong Tang, Li Xu, Xijie Guo
KEYWORDS:
Bacmid; Expression; Microsporidia; Nosema bombycis; Spore Wall Protein
JOURNAL NAME:
Agricultural Sciences,
Vol.4 No.2,
February
27,
2013
ABSTRACT: The microsporidian spore wall proteins, as the main
components of the spore wall, play a key role in spore adherence to host cells
and in recognition of the parasite by the host during the invasion process.
In this study, we used the Bac-to-Bac baculovirus expression system to express
the spore wall protein SWP26, fused to enhanced green fluorescent protein
(EGFP), in the silkworm BmN cell line. The SWP26
and EGFP genes were inserted into the
baculovirus transfer vector pFastBac1. The transfer vector pFastBac1-swp26-egfp
was transformed into the bacterium Escherichia coli DHl0Bac/Bombyx mori nucleopolyhedrovirus
(BmNPV) to construct the recombinant vBmswp26-egfp bacmid. The vBmswp26-egfp bacmid DNA was then used to transfect BmN cells to obtain the recombinant
baculovirus. Western blotting analysis of total protein lysates in BmN cells
infected by the recombinant virus showed a protein band of approximately 51
kDa, which corresponded to the deduced molecular weight of the swp26-egfp
fusion protein. In addition, a fluorescence signal was observed in the cytoplasm
and nucleoplasm of
transfected cells, indicating that SWP26 had been
successfully expressed in BmN cells. The SWP26 expression system established
in this study lays the foundation for additional molecular and cellular
studies, especially those focused on the interaction between the SWP26
protein of Nosema bombycis and the
proteins of the silkworm, Bombyx mori.