摘要
核酶（Ｒｉｂｏｚｙｍｅ）是一类具有核酸内切酶活性的ＲＮＡ分子，可特异地切割靶ＲＮＡ序列。核酶的催化中心有相对固定的序列，而切割特异性则由催化中心两侧序列与何种靶分子序列互补而决定。根据核酶这一特性，可以人工设计针对某一病毒ＲＮＡ的核酶分子，破坏病毒转...
A hammerhead ribozyme gene was synthesized directed against the sites of 110, 112 and 132 nucleotides of HBV PreS2 gene. The target gene fragment was cut from HBV-genome-containing plasmid pCP10. Both ribozyme and target gene fragments were cloned into pGEM3Zf(-)plasmid and sequenced by dideoxy chain termination method. Transcription of both gene fragments was performed in vitro utilizing T7 RNA promoter in pGEM3Zf(-)plasmid. The cleavage activity of ribozyme to substrate was observed. For further evaluating the function of riboyzme in vivo, two ribozyme-retroviral recombinant plasmids of different promoter type pDOR-ripc and tRNA-ripc were constructed with pDORneo and N2A-tRNA as vectors respectively. Recombinant vectors were transfected into viral packaging cell line PA317 introduced by lipofectamine. Recombinant retrovirus was obtained at 10 5 titer level.
出处
《病毒学报》
CAS
CSCD
北大核心
1998年第4期365-369,共5页
Chinese Journal of Virology
