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以质粒（ｓＳＶＬＤ３）为模板，通过聚合酶链反应（ＰＣＲ）扩增得到一条１３９ｂｐ的片段，它含有丁型肝炎病毒（ＨＤＶ）基因组ＲＮＡ中核酶（ｒｉｂｏｚｙｍｅ）区的ｃＤＮＡ，该核酶具有自身裂解功能，将上述片段插入到ｐＧＥＭ－３Ｚ中，经筛选、鉴定，得到一重组质粒（ｐＨＤＶ１０８），经测序发现有２个碱基变异，以该质粒为模板，通过Ｔ７ＲＮＡ聚合酶，转录出核酶的前体，并观察到其自身裂解产物，自裂率达７１％。针对核酶两个重要的单链区，设计并合成二条反义寡核背酸（ＡＳＯＮ），当在转录反应中同时加入ＡＳＯＮ后，核酶的自裂率下降均较明显，当ＡＳＯＮ浓度为１６ｕｍｏ１／Ｌ时，核酶自裂抑制率均在７０％以上。提示，ＡＳＯＮ可与核酶两个重要的单链区结合，从而抑制核酶的自裂活性。
Using the plasmid (sSVLD3) as a template, a 139 bp fragment was amplified by polymerase chain reaction (PCR), which contained the cDNA of the ribozyme region in the genomic RNA of hepatitis D virus (HDV) The above fragment was inserted into pGEM-3Z and screened and identified to obtain a recombinant plasmid (pHDV108). Two nucleotide mutations were found by sequencing. The plasmid was used as a template to transcribe the ribozyme through T7 RNA polymerase Of the precursor, and observed its own lysis products, self-cracking rate of 71%. Two antisense oligonucleotides (ASON) were designed and synthesized for two important single-stranded regions of ribozyme. When ASON was added into the transcription reaction simultaneously, the decrease of ribozyme cleavage rate was obvious. When ASON concentration was 16umo1 / L, the rate of ribozyme self-cleavage were more than 70%. It is suggested that ASON can bind to two important single-stranded regions of ribozyme to inhibit the ribozyme’s self-cleavage activity.