Identification and characterization of a novel calreticulin involved in the immune response of the Zhikong scallop, Chlamys farreri.
Calreticulin (CRT) is a multifunctional calcium-binding chaperone shared among vertebrates and invertebrates. In this study, a novel CRT (CfCRT) was identified in the Zhikong scallop Chlamys farreri by rapid amplification of cDNA ends. The full-length cDNA was composed of 1345 bp, which included a 1158 bp open reading frame, a 25 bp 5'-untranslated region (UTR) and a 162 bp 3'-UTR. The predicted molecular mass of CfCRT was 44.8 kDa. CfCRT contained three highly conserved domains (N-, P- and C-domains) essential to the function of CRT. BLAST analysis revealed significant sequence similarity (73%-92%) with CRT proteins from other mollusks. The mRNA transcripts of CfCRT were present in all the tested tissues of Zhikong scallops, with the higher expression level in the hemocytes and mantle. After stimulation by Vibrio anguillarum, the mRNA transcript of CfCRT in hemocytes was significantly upregulated. Recombinant plasmid pBCRT was successfully expressed in Escherichia coli BL21 (DE3). The recombinant (r)CfCRT protein could bind to the surface of several bacteria including the Gram-negative bacteria V. anguillarum, E. coli, and the Gram-positive bacterium Staphylococcus aureus. Moreover, rCfCRT was able to suppress their growth significantly. These results indicate that CfCRT might act as an immune effector in Zhikong scallop innate immunity.
School of Marine Science, Ningbo University, Ningbo, 315211, China; Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China.
Guanghua
G
Wang
Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, 266071, China; Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071, China.
Zengjie
Z
Jiang
Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China.
Ning
N
Yang
School of Marine Science, Ningbo University, Ningbo, 315211, China. Electronic address: zhudongfa@nbu.edu.cn.
Dongfa
D
Zhu
Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China.
Min
M
Zhang
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D037282:Calreticulin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000502:physiology*; D006434:Hemocytes; D007113:Immunity, Innate; D049897:Pectinidae / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013211:Staphylococcus aureus / Q000502:physiology*; D014733:Vibrio / Q000502:physiology*
D037282:Calreticulin; D018076:DNA, Complementary; D012333:RNA, Messenger
Bacteria-binding; Calreticulin; Chlamys farreri; Innate immunity