抗HTNV mAb 3G1单链抗体基因转化植物的研究

目的 构建抗HTNV mAb 3G1 scFv的转基因拟南芥植株.方法 从含有3G1 scFv基因的重组质粒中酶切获得含有该目的基因的表达框,将其克隆入载体pCAMBIA2301,构建植物表达载体3G1scFv-pCAMBIA2301.通过农杆菌介导的花粉管法将其转入拟南芥,PCR和Southern blotting检测是否获得转基因植株.组织化学染色检测标记基因GUS是否表达.结果 限制性内切酶酶切鉴定结果证明3G1 scFv基因被成功克隆入植物表达载体pCAMBIA2301,构建获得3G1scFv-pCAMBIA2301重组质粒.PCR和Southern blotting检测证明获得抗HTNV mAb 3G1 scFv的转基因拟南芥植株.组织化学染色结果表明,标记基因亦高效表达.结论 成功地将外源基因转入拟南芥中并表达,为进一步研究利用植物表达医用抗体奠定了基础.

Transformation of single chain Fv gene of mAb 3G1 against Hantaan virus into Arabidopsis thaliana

Object To construct the transgenic Arabidopsis thaliana containing single chain Fv gene of monoclonal antibody 3G1 against Hantaan virus. Methods The expression frame containing 3G1 scFv gene fragment was digested from recombinant 3G1 scFv-pBI121 and subeloned into pCAMBIA2301 to generate plant expression vector 3GlscFv-pCAMBIA2301. The Arabidopsis thaliana was transformed by the infdtration mediated with Agrobacterium tumefaciens. The obtained transgenic plants were analyzed by PCR, Southern blotting, and Gus staining. Results The recombinant 3GlscFv-pCAMBIA2301 was proved to be constructed suecessfully by restriction enzyme analysis. PCR and Southern blotting analysis demonstrated the integration of 3G1 scFv gene into the transgenic Arabidopsis thaliana genome. The results of Gus staining showed that the gene marker had been expressed in transformed Arabidopsis thaliana. Conclusion The foreign gene is transformed into Arabidopsis thaliana successfiuly, which lay a foundation for further study on expression of certain antibody in plant.