Davis V S, Boyle J A
College of Veterinary Medicine, Mississippi State University, Mississippi State 39762.
Avian Dis. 1990 Apr-Jun;34(2):329-35.
Viruses from three commercially available modified-live infectious bursal disease virus vaccines were propagated in tissue culture. Following this, a series of 32P-labeled probes was generated using the entire RNA genome as template for formation of randomly primed cDNAs. These probes were tested against dot blots of the three vaccine strains, as well as the USDA standard challenge strain and one field-origin strain. Dot blots were made of both crude tissue extract and LiCl-precipitated RNA genome. All three probes detected the standard challenge and field strains. Although differences in probe binding could be quantified among the strains, cross-hybridization indicated considerable homology within genomic regions preferentially transcribed under the experimental conditions.
从三种市售的传染性法氏囊病病毒活疫苗中提取的病毒在组织培养中进行繁殖。之后,以整个RNA基因组为模板,通过随机引物法合成cDNA,生成了一系列32P标记的探针。这些探针用于检测三种疫苗毒株、美国农业部标准攻击毒株和一株野外分离毒株的斑点印迹。斑点印迹采用粗组织提取物和LiCl沉淀的RNA基因组制作。所有三种探针均能检测到标准攻击毒株和野外分离毒株。虽然各毒株间探针结合的差异可以量化,但交叉杂交表明,在实验条件下优先转录的基因组区域内存在相当程度的同源性。