【摘要】 用传染性法氏囊病病毒 (IBDV)攻击 4周龄的非免疫鸡 ,以超速离心法从感染鸡法氏囊组织中分离、纯化IBDV ,应用蛋白酶K消化和Trizol(异硫氰酸胍 -酚 -氯仿法 )处理 2种方法从纯化的IBDV中抽提dsRNA。通过低熔点琼脂糖割胶 -酚 -氯仿抽提可获得纯化的dsRNA ,研究发现应用蛋白酶K消化获得的纯化RNA产量高 ,用其作模板进行RT -PCR可扩增IBDV基因组全长cDNAA片段和B片段、VP2基因和VP2 - 4 - 3基因。更多还原

【Abstract】 week old chickens unaffected by infectious bursal disease virus (IBDV) were challenged with Shanghai strains of IBDV orally. The virion were purified through ultracentrifuge from affected bursa. A proteinase K digestion based approach and Trizol RNA extraction method were used to extract dsRNA of IBDV from purified virions. The yield of dsRNA extracted by proteinase K digestion based approach was higher than that of Trizol method. dsRNA extracted by proteinase K digestion was more suitable for the amplification of full length cDNA of IBDV by long and accurate PCR.更多还原