von Weizsäcker F, Blum H E, Wands J R
Massachusetts General Hospital, Harvard Medical School, Charlestown.
Biochem Biophys Res Commun. 1992 Dec 15;189(2):743-8. doi: 10.1016/0006-291x(92)92264-x.
Hepatitis B virus is a DNA virus which replicates asymmetrically through reverse transcription of an RNA intermediate (pregenomic RNA). As a first step toward an antiviral strategy, a single synthetic oligodeoxynucleotide coding for 3 ribozyme motifs directed against three adjacent sites within the pregenomic RNA was synthesized, cloned and transcribed in vitro. Experiments utilizing 5' and 3' end labeling of RNA demonstrated that the three ribozymes accurately cleaved hepatitis B virus substrate RNA. Cleavage efficiency was similar to that of single ribozyme constructs. These results demonstrate that hepatitis B virus RNA is susceptible to ribozyme induced cleavage and illustrate that three ribozymes were simultaneously active when transcribed from a single DNA template. The expression of multiple ribozyme motifs may offer an advantage if there is high viral target sequence variability.
乙型肝炎病毒是一种DNA病毒，它通过RNA中间体（前基因组RNA）的逆转录进行不对称复制。作为抗病毒策略的第一步，合成了一种编码针对前基因组RNA中三个相邻位点的3个核酶基序的单条合成寡脱氧核苷酸，进行了克隆并在体外转录。利用RNA的5'和3'末端标记进行的实验表明，这三种核酶能准确切割乙型肝炎病毒底物RNA。切割效率与单个核酶构建体相似。这些结果表明，乙型肝炎病毒RNA易受核酶诱导的切割作用影响，并说明当从单个DNA模板转录时，三种核酶同时具有活性。如果病毒靶序列存在高度变异性，多个核酶基序的表达可能具有优势。
