Identification and molecular characterization of a Spätzle-like protein from Chinese shrimp (Fenneropenaeus chinensis).
In invertebrates, the Toll signaling pathway is important for activation of antimicrobial peptides in the innate immune system. Activation of the Toll pathway requires binding of Toll with its ligand Spätzle. Here we described a Spätzle-like protein, designated as Fc-Spz, from hemocytes of Chinese shrimp, Fenneropenaeus chinensis. The deduced amino acid sequence of Fc-Spz shares 54% identity with Spätzle-like protein of salmon louse (Lepeophtheirus salmonis). Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), quantitative real time PCR, and Western blot analyses were carried out to analyze the expression pattern and distribution profile of Fc-Spz in shrimp after challenged with bacteria and virus. The results showed that Fc-Spz mRNA was up-regulated in all the tissues tested in shrimp injected with Vibrio anguillarum and white spot syndrome virus (WSSV). The C-terminal active Fc-Spz domain (114 residues) was expressed in Escherichia coli and purified by affinity chromatography. The recombinant Fc-Spz C-114 was injected into crayfish (Procambarus clarkii) to determine the expression levels of several antimicrobial peptide genes. The results showed that recombinant Fc-Spz C-114 could up-regulate crustin 2 expression in crayfish. These results suggest that Fc-Spz may play a role in the innate immune defence of Chinese shrimp and crayfish.
Shi
Xiu-Zhen
XZ
School of Life Sciences, Shandong University, No. 27 Shanda Nanlu, Jinan, Shandong 250100, China.
Zhang
Ran-Ran
RR
Jia
Yu-Ping
YP
Zhao
Xiao-Fan
XF
Yu
Xiao-Qiang
XQ
Wang
Jin-Xing
JX
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western / Q000662:veterinary; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D020224:Expressed Sequence Tags; D006434:Hemocytes / Q000378:metabolism / Q000382:microbiology; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology; D010802:Phylogeny; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D014733:Vibrio / Q000276:immunology
D018076:DNA, Complementary; D011506:Proteins; D012333:RNA, Messenger