摘要
从RAPD扩增的鳜鱼病毒 (SCV)核酸电泳带中回收了二个片断 ,克隆子pUC19质粒 (称为SCVE36 9和SCVE45 0 )。序列分析表明插入片段分别为 36 9bp和 45 0bp与GenBank序列没有显著的同源。根据克隆序列设计两对引物P1/ P2 和P3 /P4 ,在健康鳜鱼、病鳜以及提纯的SCV核酸中进行PCR试验。结果表明 ,P1/P2 组引物在SCV基因组中扩增出特异性核酸片段 ,可作为鳜鱼病毒PCR诊断 ,检测片段为 36 9bp。
Two RAPD fragments of Siniperca chuatsi virus (SCV) genome DNA recovered from agrose gel were inserted into plasmid pUC19 (called SCVE369 and SCVE450). The sequences reported the length of cloned fragments were 369bp and 450bp. It doesn't show significant homology sequences against to those in GenBank. According those sequence two oligonucletide primersets (P 1/P 2 and P 3/P 4) were designed and used to amplify genome DNA of SCV by the method of Polymerase Chain Reaction (PCR). The results showed primer (P 1/P 2 ) could specially amplify 369bp in size's fragment in SCV genome. Furthermore, this primer set was used to test the DNA samples including the normal mandarin fish, cultured fish naturally infected with SCV and the fish artificially infected by SCV. The 369bp in size PCR product was only obtained in the mandarin fish carried SCV. Primer (P 1/P 2 ) can been used to diagnose SCV disease of mandarin fish.
出处
《水产学报》
CAS
CSCD
北大核心
2001年第1期43-46,共4页
Journal of Fisheries of China
基金
农业部和广东省科委以及广东省自然科学基金资助! (980 90 7)
关键词
鳜
鳜鱼病毒
PCR诊断
基因克隆
序列分析
Siniperca chuatsi
Siniperca chuatsi virus
polymerase chain reaction(PCR)
rapid diagnosis
