Abstract: Four pairs of primers were designed based on the full length genomic sequence of DTMUV ZJ\|407 strain. Four overlapping cDNA fragments were amplified by RT\|PCR， which were designated as A， B， C and D， respectively. Individual fragments were assembled into pBluescript Ⅱ KS （+） （pSK） vector by In\|fusion technology， and formed the full\|length cDNA clone of DTMUV ZJ\|407 （pSK\|T7\|DTMUV）. The complete DTMUV cDNA was positioned under the control of T7 promoter elements for in vitro transcription. A SmaⅠ site was introduced immediately down stream of 3′ end of DTMUV genome for in vitro linearization. Sequence analysis of the full\|length cDNA clone showed that there were nine nucleotide mutations compared with the parental virus sequence， of which seven nucleotide mutations were silent mutations and two nucleotide transitions resulted in amino acid substitutions in non\|structural protein （NS5）. In conclusion，  The full\|length cDNA clone of DTMUV ZJ\|407 strain have been successfully constructed， and which would lay a solid foundation for further rescuing DTMUV effectively and further researching DTMUV at the molecular level.

Key words: duck Tembusu virus（DTMUV）, full length cDNA clone, infectious cDNA clone