brcaa1基因新抗原表位片段的扩增、快速表达和活性鉴定

Amplification,rapid expression of new antigen epitope of brcaa1 gene and its activity identification

目的制备包含乳腺癌新抗原表位的BRCAA1部分肽段。方法根据brcaa1基因序列与RTS技术要求,设计并合成带有His尾巴的PCR引物,利用brcaa1基因质粒作模板,利用PCR技术扩增获取相应基因片段并测序证实,然后采用RTS系统试剂盒与配套设备,快速翻译合成蛋白,利用PAGE电泳与Western Blot技术,鉴定表达蛋白的活性。结果PCR获取包含乳腺癌新抗原表位的BRCAA1基因片段;测序证实所获片段是所需的目的片段;Western Blot分析证实表达的肽段具有抗原活性。结论制备了具有活性包含乳腺癌新抗原表位的BRCAA1肽段,为进一步制备单克隆抗体与建立诊断方法奠定了基础。

[Objective] To prepare the short BRCAA1 peptide including novel antigen epitope with bioactivity. [Methods] The primers with his tag for amplifying brcaal gene fragment including novel antigen epitope were designed according to the breaal sequence, the gene fragments were obtained by polymerase chain reaction based on breaal plasmid as template, and confirmed by sequencing. The short peptides including novel antigen epitope were obtained by rapid translation system, and its antigen activity was identified by Western blotting. [Results] The gene fragments including novel antigen epitope were obtained from brcaal plasmid and confirmed by PCR and sequencing analysis. The matched short peptides were fabricated by RTS. Western blotting confirmed that the matched short peptides owned antigen activity. [Conclusion] The short peptides including novel antigen epitope with antigen activity are fabricated successfully, which lays foundation for further antibody preparation and diagnosis method establishment.