Cloning and characterization of a novel C-type lectin gene from shrimp Litopenaeus vannamei.
In invertebrates, C-type lectins play crucial roles in innate immunity responses by mediating the recognition of host cells to pathogens and clearing microinvaders, which interact with carbohydrates and function as pattern recognition receptors (PRRs). A novel C-type lectin gene (LvLec) cDNA was cloned from hemocytes of Litopenaeus vannamei by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA of LvLec was of 618 bp, consisting of a 5'-terminal untranslated region (UTR) of 60 bp and a 3'-UTR of 87 bp with a poly (A) tail. The deduced amino acid sequence of LvLec possessed all conserved features critical for the fundamental structure, such as the four cysteine residues (Cys(53), Cys(128), Cys(144), Cys(152)) involved in the formation of disulfides bridges and the potential Ca(2+)/carbohydrate-binding sites. The high similarity and the close phylogenetic relationship of LvLec shared with C-type lectins from vertebrates and invertebrates. The structural features of LvLec indicated that it was an invertebrate counterpart of the C-type lectin family. The cDNA fragment encoding the mature peptide of LvLec was recombined and expressed in Escherichia coli BL21(DE3)-pLysS. The recombinant protein (rLvLec) could agglutinate bacteria E. coli JM109 depending on Ca(2+), and the agglutination could be inhibited by mannose and EDTA. These results indicated that LvLec was a new member of C-type lectin family and involved in the immune defence response to Gram negative bacteria in Litopenaeus vannamei.
Zhang
Ying
Y
Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.
Qiu
Limei
L
Song
Linsheng
L
Zhang
Huan
H
Zhao
Jianmin
J
Wang
Lingling
L
Yu
Yundong
Y
Li
Chenghua
C
Li
Fengmei
F
Xing
Kezhi
K
Huang
Bingxin
B
D000372:Agglutination Tests; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D037181:Lectins, C-Type / Q000737:chemistry / Q000235:genetics*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000235:genetics* / Q000276:immunology*; D010802:Phylogeny; D012333:RNA, Messenger; D011994:Recombinant Proteins / Q000276:immunology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D037181:Lectins, C-Type; D012333:RNA, Messenger; D011994:Recombinant Proteins