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Cloning and characterization of an invertebrate type lysozyme from Venerupis philippinarum

文献类型： 外文期刊

作者： Zhao, Jianmin 1 ; Qiu, Lihua 2 ; Ning, Xuanxuan 3 ; Chen, Aiqin 1 ; Wu, Huifeng 1 ; Li, Chenghua 1 ;

作者机构： 1.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China

2.Chinese Acad Fishery Sci, S China Sea Fishery Res Inst, Guangzhou 510301, Guangdong, Peoples R China

3.SOA, Yantai Ocean Environm Monitoring Cent Stn, Yantai 264006, Peoples R China

关键词： Venerupis philippinarum;i-type lysozyme;Tissue expression;Time-course expression

期刊名称：COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY （ 影响因子：2.231； 五年影响因子：2.215 ）

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收录情况： SCI

摘要： Lysozymes are key proteins to invertebrates in the innate immune responses against bacterial infections and providing nutrition as digestion enzymes. In the present study, an invertebrate type lysozyme (denoted as VpLYZ) was identified from Venerupis philippinarum haemocytes by cDNA library and RACE approaches. The full-length cDNA of VpLYZ consisted of 805 nucleotides with a canonical polyadenylation signal sequence AATAAA and a polyA tail, and an open-reading frame of 558 bp encoding a polypeptide of 185 amino acids with a calculated molecular mass of 20.87 kD and theoretical pI of 8.44. The high similarity of VpLYZ with other i-type lysozymes from mollusk indicated that VpLYZ should be a new member of i-type lysozyme family. Similar to most i-type lysozymes, VpLYZ possessed all conserved features critical for the fundamental structure and function of i-type lysozymes, such as three catalytic residues (Glu19, Asn72 and Ser75) and itype specific motif CL(E/L/R/H)C(I/M)C. By semi-quantitative RT-PCR analysis, mRNA transcript of VpLYZ was found to be most abundantly expressed in the tissues of gills, hepatopancreas and haemocytes, weakly expressed in the tissues of muscle, foot and mantle. After clams were challenged by Vibrio anguillarum, the mRNA level of VpLYZ in overall haemocyte population was recorded by quantitative real-time RT-PCR. VpLYZ mRNA was down-regulated sharply from 6 h to 12 h post-infection. Then, the expression level increased to the peak at 72 h and recovered to the original level at 96 h. All these results indicated that VpLYZ was involved in the immune response against microbe infection and contributed to the clearance of bacterial pathogens.

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