鸡干扰素-α基因原核表达载体的构建及高效表达

干扰素（IFN）-α因其具有很强的抗病毒活性而备受关注。本研究根据GenBank上公布的鸡IFN-α核酸序列设计1对引物，用PCR方法扩增出鸡IFN-α基因，并将其克隆到原核表达载体pET-32a及pET-30a-DsbA上，得到重组质粒pET-32a-IFN-α及pET-30a-DsbA-IFN-α。将重组质粒转化大肠杆菌Transetta（DE3）感受态细胞，经SDS-PAGE电泳及Western blotting分析，结果表明pET-30a-DsbA-IFN-α表达量较高，表达的融合蛋白分子质量约40.4 ku，表达产物主要以包涵体形式存在，且表达的融合蛋白能与His标签单克隆抗体发生特异性反应。

Construction and High Level Expression of Prokaryotic Expression Vector of Chicken Interferon-α Gene

Interferon (IFN)-α had received much more attention for its comprehensive antiviral activity. According to the relevant sequence from GenBank, a pair of specific primers was designed for amplifying chicken IFN-α gene with PCR method. The IFN-α gene was cloned into prokaryotic expression vector pET-32a and pET-30a-DsbA and the protein’s expression was identified by SDS-PAGE. Immunity characteristic of the protein was analyzed by Western blotting. The cloned pET-30a-DsbA-IFN-α/E.coli was found to produce a 40.4 ku protein at the high level. The expression products existed mainly in the form of inclusion body. These expression products could specificly react with His-tag monoclonal antibody.