Objective To obtain recombinant tropomyosin (TM).Methods In this study, the gene sequence of a TM homologous protein is obtained from the National Center of Biotechnology Information (NCBI) database, and specific primers are designed accordingly. Subsequently, using the cDNA of Litopenaeus vannamei meat as the template, the TM coding gene sequence of L. vannamei is amplified by PCR and sequenced. Finally, the prokaryotic recombinant expression system for L. vannamei TM is established using the Escherichia coli heterologous recombinant expression vector pET29a.Results After agarose gel electrophoresis (AGE) of the RNA samples, the band is clear and bright, with no diffusion observed in the lanes. The cDNA sample displays a clear band around 300 bp, indicating successful extraction of the total RNA samples from L. vannamei meat with intact structures and subsequent reverse transcription into cDNA. The PCR results indicate a single band at 900 bp for the TM coding gene of L. vannamei. BLAST analysis of the gene sequence shows that the TM coding gene is highly homologous (99.77%) to the known TM coding gene of L. vannamei. This study further constructs the TM recombinant expression vector pET29a-TM. SDS-PAGE analysis confirms efficient expression of the target protein in the host strain, yielding a soluble recombinant TM protein band with a relative molecular weight of approximately 3.7×10⁴.Conclusion This study successfully clones the TM coding gene from L. vannamei and constructs its prokaryotic expression system, enabling the efficient production of soluble recombinant TM protein.