In this study, we cloned a novel partial cDNA of toll-like receptor(TLR) gene from total RNA of gill of living giant freshwater prawn (Macrobrachium rosenbergii) of Shanghai population by RT-PCR by using primers designed from the partial sequence from transcriptone sequencing result, then it was sequenced and bioinformatically analyzed. Sequence analysis showed that the part of TLR cDNA was 1 875 bp in length and the open reading frame encoded a deduced protein with 575 amino acids residues. The part of TLR is a transmembrane protein that contains several domains like LRR,LRR-CT, LRR-NT motif in the extracellular domain and a TIR domain in the intracellular region, the common structural features of TLRs. The results of the amino acids sequence of TIR were compared with other species, indicating a highly conserved similarity among Litopenaeus vannamei,Marsupenaeus japonicus,Fenneropenaeus chinensis,Penaeus monodon,Eriocheir sinensi and Scylla paramamosian. Besides, similar RF PCR products of TLE gene were also amplified from total RNA of other different tissues including haemocytes, hepatopancreas, muscle and androgenic gland.