Cloning and characterization of a novel C-type lectin gene from shrimp Litopenaeus vannamei

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• 作者：Ying Zhang ; Limei Qiu ; Linsheng Song ; Huan Zhang ; Jianmin Zhao ; Lingling Wang ; Yundong Yu ; Chenghua Li ; Fengmei Li ; Kezhi Xing ; Bingxin Huang
• 关键词：C-type lectin ; Gene cloning ; Recombinant expression ; Bacteria agglutination ; Litopenaeus vannamei
• 刊名：Fish & Shellfish Immunology
• 出版年：2009
• 出版时间：January 2009
• 年：2009
• 卷：26
• 期：1
• 页码：183-192
• 全文大小：900 K

文摘

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 (Cys⁵³, Cys¹²⁸, Cys¹⁴⁴, Cys¹⁵²) involved in the formation of disulfides bridges and the potential Ca²⁺/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 Ca²⁺, 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.