Biotechnology and Aquiculture Laboratory, The South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 231 Xingangxi Road, Guangzhou, 510300, People's Republic of China.
Mol Biol Rep. 2011 Aug;38(6):3751-6. doi: 10.1007/s11033-010-0490-7. Epub 2010 Nov 20.
The techniques of homology cloning and anchored PCR were used to clone the fucose-binding lectin (F-type lectin) gene from Japanese sea perch (Lateolabrax Japonicus). The full-length cDNA of sea perch F-lectin (JspFL) contained a 5' untranslated region (UTR) of 39 bp, an ORF of 933 bp encoding a polypeptide of 310 amino acids with an estimated molecular mass of 10.82 kDa and a 3' UTR of 332 bp. The searches for nucleotides and protein sequence similarities with BLAST analysis indicated that the deduced amino acid sequence of JspFL was homological to the Fucose-binding lectin in other fish species. In the JspFL deduced amino acid sequence, two tandem domains that exhibit the eel carbohydrate-recognition sequence motif were found. The temporal expressions of gene in the different tissues were measured by real-time PCR. And the mRNA expressions of the gene were constitutively expressed in tissues including spleen, head-kidney, liver, gill, and heart. The JspFL expression in spleen was different during the stimulated time point, 2 h later the expression level became up-regulated, and 6 h later the expression level became down-regulated. The result indicated that JspFL was constitutive and inducible expressed and could play a critical role in the host-pathogen interaction.
采用同源克隆和锚定 PCR 技术,从日本鲈鱼(Lateolabrax japonicus)中克隆出岩藻糖结合凝集素(F 型凝集素)基因。鲈鱼 F 型凝集素(JspFL)的全长 cDNA 包含 39 bp 的 5'非翻译区(UTR)、933 bp 的开放阅读框(ORF),编码一个由 310 个氨基酸组成的多肽,预计分子量为 10.82 kDa,3'UTR 为 332 bp。核苷酸和蛋白质序列相似性的 BLAST 分析表明,JspFL 的推导氨基酸序列与其他鱼类的岩藻糖结合凝集素有同源性。在 JspFL 推导的氨基酸序列中,发现了两个串联结构域,它们表现出鳗鲡碳水化合物识别序列基序。通过实时 PCR 测量基因在不同组织中的时空表达。基因的 mRNA 表达在包括脾脏、头肾、肝脏、鳃和心脏在内的组织中持续表达。基因在脾脏中的表达在刺激时间点 2 小时后上调,6 小时后下调。结果表明,JspFL 是组成型和诱导型表达的,可能在宿主-病原体相互作用中发挥关键作用。