透明颤菌血红蛋白基因表达载体构建及转化地被菊研究

Construction of Vitreoscilla Hemoglobin Gene Expression Vector and Its Transformation in Ground-cover Chrysanthemum

根据透明颤菌(Vitreoscilla)血红蛋白基因Vgb序列设计引物,采用PCR法从pTPV-Vgb载体上克隆Vgb片段并添加酶切位点XhoⅠ,构建植物表达载体pCAMBIA1301-Pmi-Vgb。通过农杆菌介导法转化地被菊品种‘粉地毯’叶片,期望获得耐水湿能力提高的安全转基因地被菊植株。经过共培养、筛选、分化等步骤共获得了32个抗性芽。PCR检测结果表明8株呈阳性,氯酚红试验表明标记磷酸甘露糖异构酶基因(Pmi)在转基因株系中均有表达。该结果为进一步研究Vgb基因在地被菊耐水湿方面的作用奠定了基础。

Primers were designed according to the Vgb （Vitreoscilla hemoglobin）gene sequence. The Vgb sequence which added Xho I restriction sites were cloned from pTPV-Vgb vector by PCR technique. Then plant expression vector pCAMBIA1301-Pmi-Vgb was constructed. The leaves of ground-cover chrysanthe- mum cuhivar ＇Fenditan＇ were transformed through Agrobacteriurn turnefacien ,expecting to get the trans- genic chrysanthemum plants with improved resistance to waterlogging. After incubating, screening and dif- ferentiation,we got 32 resistant buds. The PCR results showed that 8 of 32 plants were positive. PCR assay showed that the Pmi gene was expressed in transgenic plants. The results laid the foundation of further re- search on the effect of Vgb gene in improving chrysanthemum＇s resistance to waterlogging.