A型禽偏肺病毒F蛋白的原核表达与抗原性分析

Prokaryotic Expression of Subgroup A Avian Metapneumovirus Virus F Protein and Its Antigenicity Analysis

利用大肠杆菌表达A型禽偏肺病毒(aMPV)F融合蛋白,并检测其抗原活性。通过扩增aMPV F融合蛋白的编码基因,与原核表达载体pBCX(pet43.1改造载体)连接构建重组表达载体pBCX/aMPV/A/F,从上清中非变性纯化重组蛋白,并进行Western-blotting分析。结果表明,该融合蛋白相对分子质量为74 kDa,具有与aMPV单克隆抗体良好的结合能力,说明此融合蛋白具有良好的抗原性,可用于抗体检测。

To express subgroup A avian metapneumovirus virus （aMPV） fusion （F） protein by Escherichia coli prokaryotic expression system and to further analyze the antigenicity of the recombinant protein, the F gene was amplified by PCR and cloned into Escherichia coli expression vector PBCX to obtain the recombinant expression vector PBCX/aMPV/A/F. The recombinant F protein was non-degeneratively purified from the supernatants and analyzed by WesternBlot.The result demonstrated the presence of a specific and simple band with a size of about 74 kDa was examined by Westem Blot using a monoclonal antibody raised against subgroup A aMPV F protein. This study indicates that the aMPV F protein can be effectively expressed in E. eoli and has a good antigenicity, Thus it can be used as an antigen for diagnosis and detection of aMPV infection.