Identification of adhesin-like protein ALP41 from Spiroplasma eriocheiris and induction immune response of Eriocheir sinensis

Spiroplasma eriocheiris is a causative agent of the tremor disease (TD) of Chinese mitten crab Eriocheir sinensis which is a novel pathogen of aquatic animals found in recent years. A gene, adhesin-like protein (ALP41), of S. eriocheiris from E. sinensis was identified and its characteristics were analyzed in present paper. The role of this pathogen’s host-binding protein in promoting immune responses was characterized through analyzing the interaction between S. eriocheiris and E. sinensis. The full-length DNA of ALP41 is 1074 bp and encodes 357 amino acid residues. The theoretical molecular weight and isoelectric point for the ALP41 are 40.94 kDa and 4.79, respectively. Since UGA is read as a tryptophan codon and not as a termination signal in most Mollicute species, the ALP41 gene was site-mutated from TGA to TGG and transcribed in Escherichia coli to full expression; the titer of rabbits anti-ALP41 serum was about 1:6000. A specific immunoreactive band was identified when rabbits anti-S. eriocheiris serum was opposed to the recombinant protein. The ALP41 band was detected using anti-ALP41 serum and the total proteins of S. eriocheiris. Realtime-PCR was used for detection of expression levels of the immune genes in E. sinensis. Among the examined genes, the mRNA expression of anti-lipopolysaccharide factor (ALF), prophenoloxidase (proPO), peroxiredoxin 6 (Prx6) and pacifastin light chain (PLC) in E. sinensis were significantly induced after ALP41 treatment.