【摘要】 利用氯化艳一蔗糖超速离心法纯化了细胞培养的鸡传染性法氏囊病（ＣＪ一８０１株）。经蛋白酶Ｋ消化，酚抽提得到的基因组ＲＮＡ在ＡＭＶ反转录酶催化下合成双链ｃＤＮＡ，通过Ｓｅｐｈａｃｒｙｌ４００柱层析得到４００ｂｎ以上的大片段。将其重组到载体质粒ｎＵＣＩ９的ＳｍａⅠ酶切位点上，转化大肠杆菌ＮＭ５２２株。通过α－互补鉴定重组体克隆，从而构建了ＩＢＤＶ的ｃＤ－ＮＡ文库。从文库中筛选出三个重组子作为用于ＩＢＤＶ诊断的探针，其中一个重组子通过序列分析加以确证，证实其定位于ＶＰ４的近３’端，并用３２Ｐ标记，进行斑点杂交，初步证实了此探针可用于鉴别ＩＢＤＶ与其他禽传染性病毒。更多还原

【Abstract】 Cj-80l strain of chicken infectious bursal disease virus was purified using the method ofCsCl-sucrose ultracentrifugation to extract the genomic RNA that was reverse transcripted to cDNA by thecatalysis of AMV reverse transcriptase, Synthesized cDNA was recombined with plasrnid pUC19 andtransformed into NM4522 strain of E. coli so a cDNA librarv was contructed. From this library three re-combinants were selected as probes to detect the existance of IBDV. Dot blott hybridization was used todetermine the IBDV form the infected cell and tissue with one of these cloned probes.The usage of nucleicacid probe was evaluated as a sirnplified means for detection of IBDV.更多还原