›› 2008, Vol. 51 ›› Issue (5): 486-491.

• RESEARCH PAPERS • Previous Articles     Next Articles

Characterization of the nonstructural gene promoter of Periplaneta fuliginosa densovirus

YANG Bo   

  • Online:2010-07-27 Published:2008-05-20


To understand the mechanism of transcriptional regulation of Periplaneta fuliginosa densovirus (PfDNV) nonstructural genes5-flanking sequence of ns3 gene (325 bp), which encompasses the region from the

5-terminus of the viral genome to the translation init iation codon of the ns3 gene, was cloned. This construction was subsequently used as a template for generation of six 5-or 3-end unidirectional truncation mutants and two site directed mutants in the PCR amplification. These amplified products were subcloned into the report plasmid pGL3-basic and the resultant plasmids were transfected into several insect cell lines. The results indicated that the 325 bp DNA sequence possessed promoter activity in insect cell lines including Sf9, Ld652, Tn368, and S2 cells. Subsequent promoter deletion analysis showed that the promoter had a TATA dependent and TATAindependent transcriptional activity. In addition, we found that the tr a nscriptional activity of this promoter could be transactivated by the viral nons tructural protein NS1 in a concentration dependent fashion in S2 cells.

Key words: Periplaneta fuliginosa densov irus (PfDNV), promoter, transactivation, insect cells, expression vector