Abstract: In order to clone and analyze the prokaryotic expression of thymosin gene from Musca domestica, the thymosin gene which was isolated from Musca domestica cDNA library, was analyzed by the bioinformatics methods in the following aspects, including general physical and chemical properties, signal peptide and subcellular localization. The expression construct pET-28a（+）-THY was preformed. The open reading frame of the thymosin gene was 384 bp that encoded a putative protein with 127 amino acids. The protein with predicted molecular weight 14.3 kD and pI of 5.22, has the conserved thymosin domain that belongs to thymosin family. The result showed that the recombinant prokaryotic expression vector pET-28a （+）-THY was successfully constructed and fusion protein was expressed in E. coli. SDS-PAGE and Western blot analysis indicated that the fusion protein that purified using Ni²⁺ affinity chromatography had the predicted size.

Key words: Musca domestica, thymosin, cloning, prokaryotic expression