文章摘要
一种凡纳滨对虾新的C型凝集素基因(LvLc2)的克隆及免疫应答特征
Molecular Cloning and characterization in immune response of a novel C-type lectin gene (LvLc2) from Litopenaeus vannamei
投稿时间:2021-01-16  修订日期:2021-03-12
DOI:
中文关键词: 凡纳滨对虾  C形凝集素  基因表达  病原刺激  天然免疫应答
英文关键词: Litopenaeus vannamei  C-type lectin  gene expression  pathogenic stimulation  innate immune response.
基金项目:天津市应用基础与前沿技术研究计划(19JCYBJC29700, 15JCZDJC33800),天津市水产生态及养殖重点实验室开放基金(TJAE2015005),国家自然科学基金(31472299)
作者单位邮编
郝少彦 天津市动植物抗性重点实验室天津师范大学生命科学学院 300387
尹丝雨 天津市动植物抗性重点实验室天津师范大学生命科学学院 
龚晨 天津市动植物抗性重点实验室天津师范大学生命科学学院 
张亦陈 天津市动植物抗性重点实验室天津师范大学生命科学学院 
耿绪云 天津市水产研究所 
刘逸尘 天津市动植物抗性重点实验室天津师范大学生命科学学院 300387
孙金生 天津市动植物抗性重点实验室天津师范大学生命科学学院 
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中文摘要:
      C型凝集素作为模式识别受体及终端效应物,同时参与“非己”识别和对入侵病原的清除过程,在甲壳动物天然免疫防御机制中至关重要。本研究根据实验室前期转录组结果提示,在凡纳滨对虾血细胞中克隆获得了一个新的C型凝集素基因(LvLc2 GenBank注册号:KR020738)。生物信息学分析显示,LvLc2基因的ORF区全长465 bp,编码154个氨基酸,5'-UTR 为11 bp,3'-UTR 为126 bp;其预测的分子量为17.16 ku,理论等电点为4.54;其推断的氨基酸序列氨基端有含17个氨基酸的信号肽序列,羧基端含有一个保守的糖识别结构域(CRD)。该基因在凡纳滨对虾各种组织中有不同程度的表达,其中在肠道中表达量最高,其次为胃、肝胰腺、血细胞等组织或细胞,在肌肉中表达量最低。利用脂多糖(LPS)和肽聚糖(PGN)刺激对虾后,比较分析了LvLc2的表达变化特征,不同类型的病原表面多糖可在刺激后不同时期引发LvLc2基因的应答改变,表明其具有潜在的广谱应答模式,同时针对不同病原又具有不同的表达特征,显示了其识别结合和间接免疫效应的差异。对其作用特征的探讨有助于更好的了解对虾C型凝集素在病原感染过程中功能和作用机制。
英文摘要:
      Litopenaeus vannamei is the largest economic shrimp breed in the world at present. However, with the expansion of shrimp culture, diseases and other problems are exposed gradually. Shrimp culture industry was affected seriously by diseases all over the world. Shrimp belongs to invertebrate, and has no typical acquired immune system. They mainly rely on the innate immune system for defense and protection when pathogens invade. In innate immune response, crustaceans recognize non-self components through pattern recognition receptors (PRRs). The components recognized by PRRs are mainly constitutive and conserved polysaccharide molecules on the surface of microorganisms, which are not produced by eukaryotic cell hosts and are necessary for the survival of pathogenic microorganisms. They are called pathogen associated molecular patterns (PAMPs). The recognition and binding of PRRs to PAMPs is the beginning of humoral immunity. The recognition of PRRS to pathogens can directly or indirectly activate proteases, hemocytes and intracellular signal pathways related to humoral immunity. A variety of immune defense reactions were triggered. It produce antibacterial substances and effective molecules to inhibit and eliminate pathogens. As a pattern recognition receptor and terminal effector, C-type lectin participates in the process of “non self” recognition and elimination of invasive pathogens, which is crucial in the innate immune defense mechanism of crustaceans. In this study, a novel C-type lectin gene (LvLc2, GenBank accession number: KR020738) was cloned from hemocytes of Litopenaeus vannamei according to our previous transcriptome results. Bioinformatics analysis showed that the ORF region of LvLc2 gene was 465 bp encoding 154 aa with 11 bp 5'-UTR and 126 bp 3'-UTR. The predicted molecular weight was 17.16 ku and the theoretical isoelectric point was 4.54. The deduced amino acid sequence contained a signal peptide sequence with 17 amino acids at the amino end and a conserved carbohydrate recognition domain (CRD) at the carboxyl end. LvLc2 gene was expressed in different tissues of L. vannamei. The main expression was in intestine, followed by stomach, hepatopancrease, hemocytes, and the lowest expression level was in muscle. After shrimps were challenged by LPS and PGN, the expression profiles of LvLc2 gene were analyzed by qPCR. Different pathogen-associated molecular patterns (PAMP) could induce response changes of LvLc2 gene after stimulation, which indicated that it had a potential broad-spectrum response pattern. At the same time, it had different expression profiles for different pathogens, which showed the differences of recognition, combination and indirect immune effects. The research of its characterization and function are helpful to better understand the mechanism of shrimp C-type lectin during the process of pathogen infection.
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