Preliminary study on the inhibition of HBV PreS2 Gene expression by retroviral mediated ribozyme transfer
Xu Dongping,Han Fenglian,Shi Hong,et al.Biological Engineering Laboratory,Institute of Infectious Diseases,Beijing,100039
In this paper, a multi-target hammerhead ri- bozyme gene was synthesized directed against 110, 122 and 132 sites of nucleotide of HBV preS2 gene. The target gene fragment was cut from HBV-genome-con- taining plasmid pCP10. Both of the ribozyme and tar- get gene fragments were cloned into pGEM3Zf(-) plasmid and sequenced by dideoxy chain termination method. The transcription of both gene fragments was performed in vitro utilizing T7 RNA promoter in pGEM3Zf(-) plasmid. The cleavage activity of ri- bozyme to substrate was confirmed in vitro. For fur- there valuating intracellular function of ribozyme, two ribozyme-retroviral recombinant plasmids with differ- ent promoter type pDOR-ripc and tRNA-ripc were constructed. Pseudo-virus was collected through rou- tine packaging procedure and transduced into 2. 2. 15 cells. RIA data showed a stable inhibition of pHSA-R antigen expression to the lowest extent of 41. 01%± 4. 16 and the highest extent of 51. 45%±4. 57 within 4 weeks after transduction. No influence, however, on HBsAg and HBeAg expression was demonstrated after ribozyme gene transfer.
【CateGory Index】： R346