Li Hongrui, Yang Jing, Chen Yubin, Guan Lili, Du LinNa, Guo YongXin, Wang Wenhui, Wang Lihao, Li Haiyan, Jiang Chao, Li Xiaokun
Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun 130118, China.
Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun 130118, China.
Protein Expr Purif. 2014 Nov;103:23-7. doi: 10.1016/j.pep.2014.03.007. Epub 2014 Mar 24.
The use of plants as expression systems for the production of recombinant proteins has distinct advantages, such as safety, ease, low cost and high yields. A plant binary expression vector, pOTBar-hPH20, containing an oleosin-hPH20 fusion gene and a Basta selection marker gene was constructed and introduced into Arabidopsis thaliana via the floral dip method. Transformed A. thaliana seed lines were obtained and analyzed by PCR. The PCR results indicated that oleosin-hPH20 fusion gene was integrated into the A. thaliana genome. The oleosin-hPH20 fusion protein was detected by SDS-PAGE and Western blot analysis. The oleosin-hPH20 fusion protein was expressed and had good antigenicity in the transgenic A. thaliana seeds. An enzyme assay suggested that the recombinant oleosin-hPH20 fusion protein had hyaluronidase activity.
利用植物作为生产重组蛋白的表达系统具有明显优势，如安全性高、操作简便、成本低且产量高。构建了一种植物双元表达载体pOTBar-hPH20，其含有油质蛋白-hPH20融合基因和Basta选择标记基因，并通过花浸染法导入拟南芥。获得了转化的拟南芥种子系并通过PCR进行分析。PCR结果表明油质蛋白-hPH20融合基因已整合到拟南芥基因组中。通过SDS-PAGE和Western印迹分析检测到了油质蛋白-hPH20融合蛋白。油质蛋白-hPH20融合蛋白在转基因拟南芥种子中表达且具有良好的抗原性。酶活性测定表明重组油质蛋白-hPH20融合蛋白具有透明质酸酶活性。
