【摘要】 根据发表的猪圆环病毒Ⅱ型(PCV2)ORF2基因序列,设计合成1对特异性引物,从分离到的PCV2吉林株(JL01)中扩增出PCV2 ORF2579 bp的核苷酸片段,克隆到表达载体pET-32a中,经BamHⅠ和HindⅢ酶切及序列分析获得阳性重组表达质粒pET-32a-ORF2。将其转化到表达宿主菌BL21中,经IPTG诱导,成功表达了ORF2基因编码的部分结构蛋白,分子量为40 kD。通过SDS-PAGE和Western检测表明,表达的重组蛋白能够被PCV2阳性血清所识别,具有良好的反应原性。更多还原

【Abstract】 Based on the published nucleotide sequence of ORF2 gene of porcine circovirus 2,a pair of primers were designed and synthesized.The 579 bp was amplified by polymerase chain reaction(PCR) from Jilin strain,and then the fragment was cloned into pET-32a vector,BamHI and HindⅢ restriction endonuclease analysis and DNA sequencing were used to identify the recombinant plasmid of pET-32a-ORF2.A recombinant vector was transformed into E.coli BL21 cells.Then about 40kD fusion protein was expressed in recombinant strain BL21 after being induced by IPTG.The expressed protein confirmed by SDS-PAGE and Western blotting analysis of reactionogenicity can react with the polyclonal antibody against PCV2.更多还原