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Acta Biochimica et Biophysica Sinica 2009 41(10):873-882; doi:10.1093/abbs/gmp079
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© The Author 2009. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

Identification of a novel negative regulatory element on the hepatitis B virus S-(+)-strand

Ying Wu1,2, Wenlu Zhang1, Yang Yang1, Bo Yu1 and Ailong Huang1,*

1 Key Laboratory of Molecular Biology on Infectious Diseases (Ministry of Education), Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400016, China
2 Key Laboratory of Chinese Medicine on Viral Diseases (Ministry of Education), Beijing University of Chinese Medicine, Beijing 100029, China

* Correspondence address. Tel: +86-23-68485230; Fax: +86-23-63822696; E-mail: ahuang{at}cqu.edu.cn


  Abstract

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.

Keywords    hepatitis B virus; plus strand; negative regulatory element; transcription

Received: February 19, 2009; Accepted: May 31, 2009
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