CAJ | 학술논문

对虾白斑综合症病毒(WSSV)编码与核酸合成代谢相关的核糖核苷酸还原酶(Ribonucleotide Reductases,RR ),与病毒 DNA 的复制有关。利用克隆技术将 RR 基因克隆到 L4440载体,构建体内合成dsRNA的大肠杆菌 HT115工程菌。诱导该工程菌合成了 RR 基因的特异双链 RNA(RR -dsRNA)和非特异双链 RNA(gfp-dsRNA),分别与 WSSV 混合共注射凡纳滨对虾,在感染72 h 后用病毒检测试剂盒提取 DNA 模板用于荧光定量 PCR 分析,结果显示 RR -dsRNA 能有效抑制 WSSV 病毒粒子的增值。
대하백반종합증병독(WSSV)편마여핵산합성대사상관적핵당핵감산환원매(Ribonucleotide Reductases,RR ),여병독 DNA 적복제유관。이용극륭기술장 RR 기인극륭도 L4440재체,구건체내합성dsRNA적대장간균 HT115공정균。유도해공정균합성료 RR 기인적특이쌍련 RNA(RR -dsRNA)화비특이쌍련 RNA(gfp-dsRNA),분별여 WSSV 혼합공주사범납빈대하,재감염72 h 후용병독검측시제합제취 DNA 모판용우형광정량 PCR 분석,결과현시 RR -dsRNA 능유효억제 WSSV 병독입자적증치。
Ribonucleotide reductases (RR )is an important enzyme during synthesis.In this research,we used RNAi technology to silence RR gene and study the effect of RR gene silence on the WSSV proliferation.We synthesized RR gene specific double-stranded RNA (RR -dsRNA)in vivo by cloning the RR gene into the L4440 plasmid and inducing dsRNA expression in E.coli HT1 15.RR -dsRNA was mixed with WSSV and injected into shrimp Litopenaeus vannamei .72 h post-Infection,WSSV DNA templates were extracted with virus detection kit for quantitative PCR analysis.The results showed that RR -dsRNA effectively inhibited WSSV proliferation.