用狄高辛探针鉴定兔出血症病毒核酸型  

Identification of RHDV Nucleic Acid's Type by DIG-labelled Probe

作　　者：吴康[1] 杜念兴[1] 徐为燕[1] 

机构地区：[1]南京农业大学,南京210095

出　　处：《农业生物技术学报》1994年第2期88-92,共5页Journal of Agricultural Biotechnology

摘　　要：RHDV人工感染病死兔肝匀浆,经氯仿处理,FEG沉淀和Sehparose-4B层析提纯病毒,再经SDS-蛋白酶K消化,酚抽提,获得纯化RHDV核酸,琼脂糖电泳,只有一条核酸带。再以狄高辛（DIG）标记2.2kbRHDV cDNA核酸探针作斑点杂交和Southern杂交,证实此带确为RHDV核酸带。用纯化病毒核酸为模板制备DIG探针与经核酸酶处理的RHDV核酸作South-ern杂交,结果显示:经RNase-Free DNase Ⅰ消化不出现杂交条带,而RNase A消化样品与RHDV核酸对照一样呈现一条明显的杂交带。结果证实RHDV核酸为DNA。Rabbit haemorrhagic disease virus (RHDV) have been purified from the liver emulasion of rabbits died of RHD. After digestion with proteinase K and extraction of purified viral preparation, the nucleic acid of high purity was isolated from virions and identified electrophoresisly. DIG -labelled 2. 2 kb fragment of RHDV DNA was hybridised with dot and southern -blot of viral nucleic acid, indicating high positive reaction. The viral nucleic acid of high purity was used as a temple for DNA DIG -labelling, and then used as a probe for hybridization. The viral nucleic acid untreated and treated with RNase - free DNase and DNase -free RNase A was electrophoresed and hybridized with DIG-lablled viral nucleic acid probe, the viral nucleic acid with RNase -free DNase digestion was undetectable in southern blot. DIG -lablled viral nucleic acid probe was also hybridized with southern blot 2. 2 kb cDNA frament. The results comfirmed that the nucleic acid of HRDV is DNA.

关 键 词：兔病 出血症 核酸探针 病毒 核酸型 

分 类 号：S858.291.5[农业科学—临床兽医学]