作　　者： ; ; ;

机构地区： 华南农业大学动物医学系

出　　处： 《畜牧兽医学报》 2002年第6期615-618,共4页

摘　　要： 依据文献报道的鸭瘟病毒 (DPV)的核苷酸序列 ,设计和合成了二对引物 ,分别为SP1和SP2 ,LP1和LP2。北京株鸭瘟病毒于鸭胚中增殖 ,差速离心和蔗糖密度梯度离心纯化 ,按酚 氯仿法抽提DNA。然后以DPVDNA为模板进行PCR ,分别扩散增出与理论相符的 42 1bp和 1 2 0 9bp的二个DNA片段 ,并将它们克隆于质粒pGEM TEasy中。经酶切和质粒PCR ,筛选含有目的基因的重组质粒。重组质粒以步移法从双方向测序 ,获 1 586bp的核苷酸序列。研究发现这 1 586bp的核苷酸序列与文献报道的UL6和UL7基因序列的同源性为 99% ,仅有一个碱基的差异。进一步作氨基酸分析 ,发现这个碱基所引起的变异为无意义突变 (CCT→CCC)。结果表明 。 According to the sequences of Duck Plague Virus(DPV)from GenBank,two pairs of primer,SP1 and SP2,LP1 and LP2,were designed.DPV was cultured in duck embryoes,and the collected allantoic fluid was treateded by differential centrifugation and sucrose density gradient centrifugations.The DNA of DPV was abstracted and was used as the model in PCR.The PCR results revealed that two DNA fragments were about 421bp and 1209bp,which were cloned into plasmid pGEM T Easy.After identification of recombinant by PCR and EcoRI digestion,the inserted DNA fragments were sequenced and sequence alignment showed 99%homology between Beijing strain and American one.The further amino acid analysis demonstrated that the mutation from CCT to CCC was no sense.

关 键 词： 鸭瘟 病毒 北京株 分子克隆 基因测序

领　　域： [农业科学] [农业科学] [农业科学]