转基因枸杞表达重组HIV壳体蛋白的鉴定

被引：6

作者：

杜国利

宋长征

张更林

孙晓光

机构：

[1] 卫生部生物技术药物重点实验室,山东省医药生物技术研究中心

来源：

中国生物制品学杂志 | 2006年 / 02期

关键词：

壳体蛋白; HIV; 转基因; 枸杞;

D O I：

10.13200/j.cjb.2006.02.28.dugl.007

中图分类号：

R346 [];

学科分类号：

摘要：

目的构建转基因枸杞表达系统,并表达HIV壳体蛋白。方法以重组质粒pET-3a-MA4-CA为模板,PCR扩增MA4-CA基因,克隆至pGEM-T质粒中,转化DH5α,经双酶切后重组于pCAMBIA1305.2载体中,构成pCAMBIA1305.2-MA4-CA载体,经农杆菌介导叶盘转化法转化枸杞,培育转基因枸杞植株,进行芽、根诱导。结果在植物培养箱中,转基因植株成功地进行了分化及生根的诱导。PCR扩增分析表明转基因植株内存在目的基因,并与启动子p35S相连。ELISA及Westernblot实验证实在转基因枸杞叶子提取物中存在壳体蛋白。结论初步建立了表达具有免疫原性的HIV壳体蛋白的转基因枸杞表达系统。

引用

页码：127 / 129+133 +133

页数：4

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