Tags

Type your tag names separated by a space and hit enter

Cloning and characterization of a novel C-type lectin from Zhikong scallop Chlamys farreri.
Mol Immunol. 2007 Feb; 44(5):722-31.MI

Abstract

C-type lectin is a family of Ca2+ dependent carbohydrate-recognition proteins which play crucial roles in the innate immunity of invertebrates by mediating the recognition of host cells to pathogens and clearing microinvaders as a pattern recognition protein (PRP). The cDNA of Zhikong scallop Chlamys farreri C-type lectin (designated CFLec-1) was cloned by expressed sequence tag (EST) and RACE techniques. The full-length cDNA of CFLec-1 was 1785 bp, consisting of a 5'-terminal untranslated region (UTR) of 66 bp and an unusually long 3' UTR of 1040 bp with seven polyadenylation signal sequences AATAAA and a poly(A) tail. The CFLec-1 cDNA encoded a polypeptide of 221 amino acids with a putative signal peptide of 15 amino acid residues and a mature protein of 206 amino acids. Analysis of the protein domain features indicated a typical long-form carbohydrate-recognition domain (CRD) of 130 residues in the CFLec-1 deduced amino acid sequence. The expression pattern of CFLec-1 transcripts in healthy and bacterial challenged scallops was studied by semi-quantitative RT-PCR. mRNA transcripts of CFLec-1 could be mainly detected in the tissues of haemocytes, gill, gonad and mantle of unchallenged scallops, whereas the expression of CFLec-1 transcripts was increased in all the tested tissues after heat-killed Vibrio anguillarum challenge. The temporal expression of CFLec-1 mRNA in haemolymph challenged by Micrococcus luteus and V. anguillarum was both up-regulated and reached the maximum level at 8 and 16 h post stimulation, respectively, and then dropped back to the original level. In order to investigate its immune functions, CFLec-1 was recombined and expressed in Escherichia coli BL21(DE3)-pLysS as a fusion protein with thioredoxin. The recombinant CFLec-1 agglutinated bacteria E. coli JM109 in vitro, and the agglutination was Ca2+ dependent which could be inhibited by EDTA. But it did not agglutinate M. luteus, Candida lipolytica and animal erythrocytes including rabbit, rat, mouse, chicken, human group A, human group B, human group O. Meanwhile, the recombinant CFLec-1 could inhibit the growth of both E. coli JM109 and M. luteus, but no inhibition activity against V. anguillarum. These result indicated that CFLec-1 was a constitutive and inducible PRP which was involved in the reorganization and clearance of invaders in scallop.

Authors+Show Affiliations

Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, PR China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16777225

Citation

Wang, Hao, et al. "Cloning and Characterization of a Novel C-type Lectin From Zhikong Scallop Chlamys Farreri." Molecular Immunology, vol. 44, no. 5, 2007, pp. 722-31.
Wang H, Song L, Li C, et al. Cloning and characterization of a novel C-type lectin from Zhikong scallop Chlamys farreri. Mol Immunol. 2007;44(5):722-31.
Wang, H., Song, L., Li, C., Zhao, J., Zhang, H., Ni, D., & Xu, W. (2007). Cloning and characterization of a novel C-type lectin from Zhikong scallop Chlamys farreri. Molecular Immunology, 44(5), 722-31.
Wang H, et al. Cloning and Characterization of a Novel C-type Lectin From Zhikong Scallop Chlamys Farreri. Mol Immunol. 2007;44(5):722-31. PubMed PMID: 16777225.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cloning and characterization of a novel C-type lectin from Zhikong scallop Chlamys farreri. AU - Wang,Hao, AU - Song,Linsheng, AU - Li,Chenghua, AU - Zhao,Jianmin, AU - Zhang,Huan, AU - Ni,Duojiao, AU - Xu,Wei, Y1 - 2006/06/14/ PY - 2006/03/25/received PY - 2006/04/24/revised PY - 2006/04/25/accepted PY - 2006/6/17/pubmed PY - 2007/3/6/medline PY - 2006/6/17/entrez SP - 722 EP - 31 JF - Molecular immunology JO - Mol. Immunol. VL - 44 IS - 5 N2 - C-type lectin is a family of Ca2+ dependent carbohydrate-recognition proteins which play crucial roles in the innate immunity of invertebrates by mediating the recognition of host cells to pathogens and clearing microinvaders as a pattern recognition protein (PRP). The cDNA of Zhikong scallop Chlamys farreri C-type lectin (designated CFLec-1) was cloned by expressed sequence tag (EST) and RACE techniques. The full-length cDNA of CFLec-1 was 1785 bp, consisting of a 5'-terminal untranslated region (UTR) of 66 bp and an unusually long 3' UTR of 1040 bp with seven polyadenylation signal sequences AATAAA and a poly(A) tail. The CFLec-1 cDNA encoded a polypeptide of 221 amino acids with a putative signal peptide of 15 amino acid residues and a mature protein of 206 amino acids. Analysis of the protein domain features indicated a typical long-form carbohydrate-recognition domain (CRD) of 130 residues in the CFLec-1 deduced amino acid sequence. The expression pattern of CFLec-1 transcripts in healthy and bacterial challenged scallops was studied by semi-quantitative RT-PCR. mRNA transcripts of CFLec-1 could be mainly detected in the tissues of haemocytes, gill, gonad and mantle of unchallenged scallops, whereas the expression of CFLec-1 transcripts was increased in all the tested tissues after heat-killed Vibrio anguillarum challenge. The temporal expression of CFLec-1 mRNA in haemolymph challenged by Micrococcus luteus and V. anguillarum was both up-regulated and reached the maximum level at 8 and 16 h post stimulation, respectively, and then dropped back to the original level. In order to investigate its immune functions, CFLec-1 was recombined and expressed in Escherichia coli BL21(DE3)-pLysS as a fusion protein with thioredoxin. The recombinant CFLec-1 agglutinated bacteria E. coli JM109 in vitro, and the agglutination was Ca2+ dependent which could be inhibited by EDTA. But it did not agglutinate M. luteus, Candida lipolytica and animal erythrocytes including rabbit, rat, mouse, chicken, human group A, human group B, human group O. Meanwhile, the recombinant CFLec-1 could inhibit the growth of both E. coli JM109 and M. luteus, but no inhibition activity against V. anguillarum. These result indicated that CFLec-1 was a constitutive and inducible PRP which was involved in the reorganization and clearance of invaders in scallop. SN - 0161-5890 UR - https://www.unboundmedicine.com/medline/citation/16777225/Cloning_and_characterization_of_a_novel_C_type_lectin_from_Zhikong_scallop_Chlamys_farreri_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0161-5890(06)00165-9 DB - PRIME DP - Unbound Medicine ER -