【摘要】：研究豬圓環(huán)病毒2型(PCV2)Cap蛋白的抗原性,為開(kāi)發(fā)PCV2的檢測(cè)方法奠定基礎(chǔ)。以PCV2CAU0673毒株的DNA為模板,擴(kuò)增獲得702bp的目的片段,擴(kuò)增產(chǎn)物克隆入pET30a(+)原核表達(dá)載體,構(gòu)建pET30a-PCV2-ORF2重組質(zhì)粒,轉(zhuǎn)入大腸埃希菌BL21(DE3);在37℃以1mmol/L IPTG誘導(dǎo)表達(dá)6h;采用Ni-NTA樹(shù)脂親和層析純化重組蛋白,并用不同濃度的尿素對(duì)純化蛋白進(jìn)行復(fù)性。SDSPAGE分析表明,該ORF2編碼基因在大腸埃希菌中得到表達(dá),蛋白大小約為34ku;Western blot檢測(cè)結(jié)果表明,該重組Cap蛋白與PCV2陽(yáng)性血清發(fā)生特異性反應(yīng),與NA-PRRSV和PPV1血清不發(fā)生交叉反應(yīng)。成功構(gòu)建了PCV2-ORF2原核表達(dá)載體,實(shí)現(xiàn)了在大腸埃希菌中的表達(dá),純化后的復(fù)性蛋白具有較好的反應(yīng)原性,為豬圓環(huán)病毒2型檢測(cè)方法建立或試劑盒的開(kāi)發(fā)奠定了基礎(chǔ)。
[Abstract]:The antigenicity of porcine circovirus type 2 (PCV2) Cap protein was studied. Using the DNA of PCV2CAU0673 strain as template, the target fragment of 702bp was amplified and cloned into pET30a () prokaryotic expression vector. The recombinant plasmid of pET30a-PCV2-ORF2 was constructed and transferred into BL21 (DE3) of Escherichia coli, and was induced by 1mmol/L IPTG for 6 h at 37 鈩，

本文編號(hào)：2331385