Original Articles
Copyright ©The Author(s) 2000. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 15, 2000; 6(3): 377-380
Published online Jun 15, 2000. doi: 10.3748/wjg.v6.i3.377
Construction of HBV-specific ribozyme and its recombinant with HDV and their cleavage activity in vitro
Shu-Juan Wen, Kai-Jun Xiang, Zhen-Hua Huang, Rong Zhou, Xue-Zhong Qi
Shu-Juan Wen, Zhen-Hua Huang, Rong Zhou and Xue-Zhong Qi, Gene Center of Nanfang Hospital, First Military Medical University, Guangzhou 510515, Guangdong Province, China
Kai-Jun Xiang, School of Life Sciences, University of Science and Technology of China, Hefei 230027, Anhui Province, China
Author contributions: All authors contributed equally to the work.
Supported by Natural Science Foundation of Guangdong Province, No. 940311.
Correspondence to: Shu Juan Wen, Gene Center of Nanfang Hospital, First Military Medical University, Guangzhou 510515, Guangdong Province, China. zhou@fimmu.edu.cn
Telephone: +86-20-85141044 Fax: +86-20-87730347
Received: January 11, 2000
Revised: February 11, 2000
Accepted: February 18, 2000
Published online: June 15, 2000
Abstract

AIM: To construct the recombinant of HDV cDNA and HBV-specific ribozyme gene by recombinant PCR in order to use HDV as a transporting vector carrying HBV-specific ribozyme into liver cells for inhibiting the replication of HBV.

METHODS: We separately cloned the ribozyme (RZ) gene and recombinant DVRZ (comprising HDV cDNA and HBV-specific ribozyme gene) into the downstream of T7 promoter of pTAdv-T vector and studied the in vitro cleavage activity of their transcripts (rRZ, rDVRZ) on target RNA (rBVCF) from in vitro transcription of HBV C gene fragment(BVCF).

RESULTS: Both the simple (rRZ) and the recombinant ribozyme rDVRZ could efficiently catalyze the cleavage of target RNA (rBVCF) under different temperatures (37 °C, 42 °C and 55 °C) and Mg2+ concentrations (10 mmol/L, 15 mmol/L and 20 mmol/L) and their catalytic activity tended to increase as the temperature was rising. But the activity of rRZ was evidently higher than that of rDVRZ.

CONCLUSION: The recombinant of HDV cDNA and ribozyme gene had the potential of being further explored and used in gene therapy of HBV infection.

Keywords: hepatitis B virus, hepatitis D virus, ribozyme gene, recombinant DVRZ