摘要
参照已知猪传染性胸膜肺炎放线杆菌血清2型菌株毒素 (Apx A)序列设计1对特异性引物,经PCR扩增得到目的片段约1096bp,然后克隆入pMD18\|T载体中,经测序比较,与标准序列的核苷酸同源性达98.5%。从T-载体中切下目的片段后,定向克隆入pET32a(+)中,转化BL21(DE3),经诱导后,SDS-PAGE结果显示,毒素 得到高效表达,Western-blotting检测呈阳性。这为研制猪传染性胸膜肺炎新型疫苗和建立有效的诊断方法奠定了基础。
The ApxⅢA gene from serotype 2 strain of Actinobacillus pleuropneumoniae was amplified by PCR with a size of 1()096 bp. The amplified DNA fragment was cloned into pMD18-T, and sequenced. The result of sequencing showed that the consistency was 98.5% compared with that of standard strains. Inserted in pET-32a, and transformed in Escherichia coli BL21 (DE3), the fragment was expressed after induced, and it could be detected by Western blotting. These results become the base of researching and producing novel vaccine and developing effective diagnostic method.
出处
《扬州大学学报(农业与生命科学版)》
CAS
CSCD
2004年第3期19-21,共3页
Journal of Yangzhou University:Agricultural and Life Science Edition
基金
江苏省创新人才基金资助项目(BK2003424)
关键词
猪
胸膜肺炎放线杆菌
毒素Ⅲ
克隆
序列分析
表达
pig
Actinobacillus pleuropneumoniae
ApxⅢA
cloning
, sequencing
expression