Identification of a novel negative regulatory element on the hepatitis B virus S-(+)-strand.
In this study, we scanned the whole hepatitis B virus (HBV) genome for the identification of potential regulatory elements located on the S-(+)-strand. With pCDNA3.1-HBV1.3 as template which contains 1.3-fold HBV whole genome, HBV fragments were amplified by PCR methods, and then inserted into the upstream of a heterologous luciferase reporter vector (pGL3control) in antisense orientation, allowing the HBV expression from the S-(+)-strand. We found that the reporter plasmid containing nt 509-1(3182)-2639 of HBV inhibited luciferase gene transcription and expression in HepG2 cells. Our results strongly suggested that nt 453-250 of HBV may act as a novel negative regulatory element, which has not been reported before. Serial deletion analyses further indicated that nt 453-250 sequence of HBV genome would be the minimal sequence essential for the inhibitory effect of the novel negative regulatory element.
Wu
Ying
Y
Key Laboratory of Molecular Biology on Infectious Diseases (Ministry of Education), Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing, China.
Zhang
Wenlu
W
Yang
Yang
Y
Yu
Bo
B
Huang
Ailong
A
D016373:DNA, Antisense / Q000494:pharmacology*; D004279:DNA, Viral / Q000032:analysis / Q000494:pharmacology; D017930:Genes, Reporter / Q000502:physiology; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D012045:Regulatory Sequences, Nucleic Acid / Q000187:drug effects* / Q000502:physiology; D012054:Reinforcement, Psychology; D014407:Tumor Cells, Cultured
D016373:DNA, Antisense; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens