Spiralin-like protein SLP31 from Spiroplasma eriocheiris as a potential antigen for immunodiagnostics of tremor disease in Chinese mitten crab Eriocheir sinensis.
Spiroplasma eriocheiris caused massive mortality of Chinese mitten crab Eriocheir sinensis but little is known about the molecular characteristics of this microorganism. We described here the identification of a spiralin-like protein (SLP31) from S. eriocheiris and expression in Escherichia coli. Analysis of the nucleotide sequence revealed that the clone has an open reading frame of 837 bp encoding a protein of 279 amino acids. Theoretical isoelectric point and molar mass for SLP31 are 7.72 and 31 kDa, respectively. The similarity of SLP31 deduced amino acid sequence shared with the spiralin from other species indicated that the gene may be a member of spiralin family. The TGA codon in Spiroplasma serves not as a stop signal but as a code for the amino acid tryptophan. After cloning the SLP31, the gene was site-mutated from TGA to TGG and transcribed in E. coli to full expression of SLP31. The purified recombinant protein was used to determine the immune reactivity by Western blotting which suggests that SLP31 could be a good antigen for immunodiagnostic of tremor disease in E. sinensis.
Meng
Qingguo
Q
Jiangsu Key Laboratory for Biodiversity & Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing, 210046, China.
Gu
Wei
W
Bi
Keran
K
Ji
Huyan
H
Wang
Wen
W
D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D001431:Bacteriological Techniques / Q000379:methods*; D001483:Base Sequence; D015153:Blotting, Western; D003386:Brachyura / Q000382:microbiology*; D002681:China; D003001:Cloning, Molecular; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D007118:Immunoassay / Q000379:methods; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013149:Spiroplasma / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; C028588:spiralin