牡蛎诺如病毒受体合成基因类FUT10的克隆与组织表达

Molecular cloning and expression of FUT10-like gene for norovirus receptor synthesis in Crassostrea gigas

中文关键词:  太平洋牡蛎  类FUT10基因  组织血型抗原  克隆  组织表达

英文关键词:Crassostrea gigas  FUT10-like gene  histo-blood group antigens  clone  mRNA expression

中文摘要:

      目的 克隆太平洋牡蛎类FUT10基因的cDNA序列, 探究类FUT10 mRNA在牡蛎各组织中的表达差异。方法 基于同源比对方法, 利用cDNA末端快速扩增(rapid amplification of cDNA ends, RACE)技术克隆完整的类FUT10基因, 通过实时荧光定量聚合酶链式反应(real-time quantitative polymerase chain reaction, qRT-PCR)分析其组织分布。结果 类FUT10基因cDNA长度为2291 bp, 含有111 bp的5’非翻译区、1401 bp编码466个氨基酸的开放阅读框及779 bp的3’非翻译区。基因进化树显示类FUT10与FUT10家族较相似。类FUT10 mRNA在太平洋牡蛎的肝胰腺中表达量高于鳃等组织。结论 本研究克隆了牡蛎类FUT10基因, 证实其在牡蛎肝胰腺中有显著表达, 推测牡蛎很可能具有与人类相似的Ⅱ型类Lewis血型抗原合成路径, 研究结果为进一步探究牡蛎富集诺如病毒的分子机制、探索诺如病毒的食源性防控策略提供基础。

英文摘要:

      Objective To clone the cDNA sequence of FUT10-like gene of Crassostrea gigas and explore the difference of FUT10-like mRNA expression in different tissues of Crassostrea gigas. Methods Based on the method of homologous alignment, the complete FUT10-like gene was cloned using rapid amplification of cDNA ends (RACE) technology, and its tissue distribution was analyzed by real-time quantitative polymerase chain reaction (qRT-PCR). Results The length of cDNA of FUT10-like gene was 2291 bp, which contains 111 bp 5’untranslated region, 1401 bp open reading frame encoding 466 amino acids and 779 bp 3’untranslated region. The genetic evolution tree showed that the FUT10-like family was similar to the FUT10 family. The expression level of FUT10-like mRNA in the hepatopancreas of Crassostrea gigas was higher than that in the gills and other tissues. Conclusion In this study, the FUT10-like gene of Crassostrea gigas is cloned, and it is confirmed that it is significantly expressed in the hepatopancreas of Crassostrea gigas. It is speculated that Crassostrea gigas probably have a type II Lewis-like blood group antigen synthesis pathway similar to that of human. The research results provide a basis for further exploring the molecular mechanism of enrichment of norovirus in Crassostrea gigas and food-borne prevention and control strategies of norovirus.