Cloning and characterization of a mannose binding C-type lectin gene from salivary gland of Aedes albopictus

Particle and Fibre Toxicology

Jiahong Wu³  Yu Sun²  Jian Liu³  Fangzhan Li³  Yu Wang¹  Jinzhi Cheng³ 

[1] Guizhou Center for Disease Control and Prevention, Guiyang, Guizhou 550004, China;Affiliated Hospital of Guiyang Medical College, Guiyang, Guizhou 550004, China;Lab for Modern Pathogen Biology, Guiyang Medical College, Guiyang, Guizhou 550004, China

关键词: Agglutinating activity;    Prokaryotic expression;    Ae. albopictus;    Salivary gland;    C-type lectin;   

Others  :  1183543
DOI  :  10.1186/1756-3305-7-337

 received in 2014-01-28, accepted in 2014-07-08,  发布年份 2014

【 摘 要 】

Background

The studies on sialomes have shown that hematophagous mosquito saliva consists of a lot of pharmacologically active proteins, in which C-type lectins have been identified and regarded as an important component of saliva. The previous studies showed that C-type lectins play crucial roles not only in innate immunity but also in promoting disease transmission in mammals. However, the function and mechanism of C-type lectins from the mosquito sialome is still elusive.

Methods

A putative C-type lectin gene (Aalb_CTL1) was cloned and expressed from Aedes albopictus by RT-PCR. The deduced amino acid sequence was analyzed by bioinformatic methods. The gene expression profiles in different tissues and various blood-fed stages of Ae. albopictus were examined by Real-Time qRT-PCR and the biological functions of the recombined mature Aalb_CTL1 were tested by hemagglutination and sugar inhibitory agglutination assays. Moreover, the capabilities of rAalb_CTL1 against microorganisms were measured by microbial-agglutination assay.

Results

The full-length Open reading frame (ORF) of Aalb_CTL1 consisted of 462 bp, encoding 153 amino acid residues. The deduced amino acid sequence contained a putative signal peptide of 19 amino acids. It also contained a CRD domain with a WND (Trp¹³⁷-Asn¹³⁸-Asp-¹³⁹) motif that needed calcium for the hemagglutinating activity and an imperfect EPS (Glu¹²⁸-Pro¹²⁹-Ser¹³⁰) motif that had a predicted ligand binding specificity for mannose. The mRNA level of Aalb_CTL1 was much higher in female mosquito salivary gland than those in fat body and midgut which was down-regulated in salivary gland after blood feeding. The rAalb_CTL1 contained not only hemagglutinating activity and a high affinity with mannose but also agglutinating activity against yeast C. albicans and Gram-positive bacteria S. aureus in Ca²⁺ dependent manner.

Conclusion

Aalb_CTL1 was a mannose-binding C-type lectin and constituted one of the important components in saliva of Ae. albopictus, which could be involved in the defense against yeast and Gram-positive bacteria infection.

【 授权许可】

   
2014 Cheng et al.; licensee BioMed Central Ltd.

【 图 表 】

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