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提取罗氏沼虾(Macrobrachium rosenbergii)上海养殖群体活体鳃、肌肉、肝胰腺、血液和雄性腺5种组织的总RNA;应用RT-PCR技术,从鳃组织中克隆到Toll样受体(TLR)基因部分cDNA序列,并进行生物信息学分析。基因序列分析表明:所得罗氏沼虾TLR基因cDNA序列长1 875 bp,包含1 728 bp的开放阅读框,可编码575个氨基酸残基;该部分TLR是一个跨膜蛋白,存在胞外区LRR、LRR-CT、LRR-NT结构域及关键的胞内区TIR结构域,具备Toll样受体家族的典型结构特征;TIR结构域与凡纳滨对虾(Litopenaeus vannamei)和日本囊对虾(Marsupenaeus japonicus)、中国明对虾(Fenneropenaeus chinensis)、斑节对虾(Penaeus monodon)、中华绒螯蟹(Eriocheir sinensis)、拟穴青蟹(Scylla paramamosian)Toll样受体基因TIR区域的氨基酸序列具有很高的相似性。
Total RNA was extracted from 5 tissues of gavage, muscle, hepatopancreas, blood and gonads of Macrobrachium rosenbergii cultured in Shanghai. Toll-like receptor (TLR) gene was cloned from gill tissue by RT-PCR cDNA sequence, and bioinformatics analysis. The sequence analysis showed that the cDNA sequence of TLR gene of M. rosenbergii was 1 875 bp in length and contained a 1 728 bp open reading frame encoding 575 amino acid residues. This partial TLR is a transmembrane protein with extracellular LRR, LRR-CT, LRR-NT domain and key intracellular region TIR domain, which have the typical structural features of Toll-like receptor family; TIR domain and Litopenaeus vannamei and Japanese shrimp (Marsupenaeus japonicus) , The amino acid sequences of the TIR region of Toll-like receptor genes in Fenneropenaeus chinensis, Penaeus monodon, Eriocheir sinensis and Scylla paramamosian have high similarity .