【摘要】 参照国外已发表的M41高可变区S1序列设计一对引物 ,跨度为 1 .7kb左右。采用差速离心法浓缩病毒 ,提取病毒RNA ,经RT_PCR扩增 ,得到与设计同样大小的PCRDNA片段 ,将PCR产物纯化 ,与质粒载体 (pGEM_TEasyVector)连接 ,并转入大肠杆菌JM1 0 9,获取阳性克隆。增菌培养后用小量碱提取法提取质粒 ,利用PCR扩增法与EcoRI酶切分析进行鉴定。对阳性质粒DNA进行测序 ,利用Omiga2 .0、Dnasis等分子生物学软件进行分析 ,并与标准M41株、T株等进行序列比较 ,结果表明 :山东传支A株与标准M41株同源性较高 ,最大同源率为 99% ;与标准T株同源性较差 ,最大同源率为 80 %。理论酶切图谱与M41相同 ,为同一基因型。更多还原

【Abstract】 The S 1 gene hypervariable region primer specific for IBV_M 41 stain was designed and a fragment of 1.7 kb to IBV_A strain was gotten by reverse transcription_polymerase chain reaction.The amplified product was purified and inserted into PGEMR_T Easy vector,then transformed into JM109 bactera.In the same time,the positive plasmid DNA by restriction enzyme assay was sequenced and compared with other IBV strain such as M 41 ,T ect.using OMIGA 2.0 and DNASIS software.Then the results showed the homogeneity of IBV_A strain with IBV_M 41 strain was 99 %,with IBV_T staint was 80 %.The acadermic enzyme cleavage fingerprinting atlas showed the IBV_A strain was the same genetype with IBV_M 41 and partially common antigen with T strain.It’s the same results with the serotype.更多还原