Cloning, Expression of apxl Gene of Actinobacillus pleuropneumoniae and Development of ELISA
Cloning, Expression of apxl Gene of Actinobacillus pleuropneumoniae and Development of ELISA
复制标题
DOI：
--
复制
发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Tang Xian-chun
中科院分区：
文献类型：
--
作者：
Tang Xian-chun
Based on the published nucleotide sequence of the apxICA of Actinobacillus pleuropneumoniae in Genbank(S4074), a pair of primers were designed. A 3 640 bp(4 687 - 8 326 bp)gene fragment was amplified by PCR from the isolated strain of A. pleuropneumoniae serovar 1. Then, it was cloned into pMD18-T, identified by both restriction endonuclease and sequence analysis, and inserted into pET-28a expression vector to yield the expression plasmid. SDS-PAGE result indicated expression of apxICA in BL21 (DE3), Western blot analysis showed the protein's immunogenicity. Using the expressed protein, ELISA was established to detect serum antibody against ApxI. The feature of ELISA to detect highly virulent A. pleuropneumoniae strains infection was proved by primary clinical application.
