Open AccessConstruction of HBV-specific ribozyme and its recombinant with HDV and their cleavage activityin vitro
Author(s) -
Shu Juan Wen
Publication year - 2000
Publication title -
world journal of gastroenterology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.427
H-Index - 155
eISSN - 2219-2840
pISSN - 1007-9327
DOI - 10.3748/wjg.v6.i3.377
Subject(s) - ribozyme , recombinant dna , microbiology and biotechnology , complementary dna , virology , biology , mammalian cpeb3 ribozyme , rna , vs ribozyme , gene , cleavage (geology) , biochemistry , paleontology , fracture (geology)
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°, 42° and 55°) and Mg(2+) concentrations (10mmol/L, 15mmol/L and 20mmol/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.