利用人内皮抑素基因构建植物表达载体及对烟草的遗传转化

Construction of Plant Expression Vector with Human Endostatin Gene and Transformation of Tobacco

将人内皮抑素(endostatin)基因重组于植物双元表达载体pGA 643,得到重组质粒pGE.用三亲融合法将其导入农杆菌LBA 4404中,采用叶盘法转化烟草,经诱导与筛选,获得了卡那霉素抗性植株.提取抗性植株总DNA,通过PCR扩增和Sou thern杂交检测,筛选出了整合有外源基因的转化植株.为进一步研究人内皮抑素在烟草中的表达及生物学功能奠定了基础.

The plant expression vector pGE, carrying a human endostatin gene,was constructed. pGE was introduced into Agrobacterium tumefaciens by triparental mating. Tobacco leaf discs were transformed via Agrobacterium tumefaciens mediated procedure. Transgenic tobacco plantlets grew in induction medium and plants resistant to kanamycin were obtained. The PCR and Southern blotting analysis confirmed that human endostatin gene had integrated into the genome of tobacco.