摘要
为了克隆从患肺炎犊牛和健康犊牛上分离的多杀性巴氏杆菌血红素结合受体(heme acquisition system receptor,HasR)基因并进行序列分析,分别以致犊牛肺炎和健康犊牛携带的多杀性巴氏杆菌DNA为模板,通过PCR反应扩增出血红素结合受体基因的全部序列,PCR产物纯化后克隆到pMD19-T载体上,经菌液PCR和酶切鉴定后进行序列测定。各菌株HasR基因序列比对结果发现,Pm-BS-a、Pm142-x-4、Pm142-x-3、Pm149-x、Pm-BS-d的序列之间亲源关系较近,而Pm149-xby与参考菌株Pm70的同源性较高。健康牛源的多杀性巴氏杆菌与致犊牛肺炎多杀性巴氏杆菌之间HasR基因的同源性非常高,说明致犊牛肺炎多杀性巴氏杆菌是一种条件性致病菌,其引起的犊牛肺炎可能属内源性感染。
To clone and do the sequence analysis of the heme acquisition system receptor of Pasteurella multocida from healthy calves and calves with pneumonia,all DNA fragment of HasR gene was amplified from the chromosomal DNA of Pasteurella multocida from healthy calves and calves with pneumonia by PCR and cloned into pMD19-T vector and the cloned fragment was sequenced after identified by PCR and two restriction enzyme digestion.DNA sequence comparison of the amplified fragment with the published sequence of HasR gene in GenBank（strain Pm70） showed high nucleotide homology among the Pm-BS-a,Pm142-x-4,Pm142-x-3,Pm149-x,Pm-BS-d,and there was a higher homology between Pm149-x and reference strain Pm70.The high homology of HasR gene between healthy calves and calves with pneumonia indicated that the Pasteurella multocida inducing pneumonia of calf might be come from upper respiratory of their own or their parents.
出处
《中国畜牧兽医》
CAS
北大核心
2011年第8期56-59,共4页
China Animal Husbandry & Veterinary Medicine
基金
牛羊疫病防治集成与示范(2007ZX02)
关键词
多杀性巴氏杆菌
血红素获得受体
克隆
序列分析
Pasteurella multocida
heme acquisition system receptor
cloning
sequence analysis
