Abstract: The aim of this study is to clone the cDNA sequence of Musca domestica 14-3-3（MD14-3-3），analysis the gene and encoded protein sequnece of MD14-3-3 by the bioinformatics methods in the following aspects, such as general physical and chemical properties, hydrophobicity, signal peptide, secondary structure and subcellular localization. Results showed that the open reading frame of the MD14-3-3 was 771 bp that encoded a putative protein with 257 amino acids. The protein, with predicted molecular weight 29.35 kD and pI of 5.78, had one active site of family 14-3-3, without signal peptide. It was a hydrophilicity acidic protein which was mainly located in cell nucleus possible, containing many potential modified sites. The secondary structures were mainly composed of αcoiling and random coil. The gene coding for MD14-3-3 was amplified by polymerase chain reaction（PCR）, then the PCR product was transformed into the E.coli DH5α through being linked with pET 28a（+）vector. As demonstrated by PCR, double enzyme digestion and DNA sequencing, it was confirmed that the recombinant expression plasmid was construction succeed. Key words: Musca domestic 14-3-3 Gene cloning Bioinformatics