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.
Key Laboratory of Molecular Biology on Infectious Diseases (Ministry of Education), Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing, China.
Ying
Y
Wu
Wenlu
W
Zhang
Yang
Y
Yang
Bo
B
Yu
Ailong
A
Huang
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