透明颤菌血红蛋白基因表达载体构建及转化地被菊研究
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国家“863”计划项目(2011AA10020804,2013AA102706)


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

    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。

    Abstract:

    Primers were designed according to the Vgb (Vitreoscilla hemoglobin)gene sequence.The Vgb sequence which added XhoⅠ 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 chrysanthemum cultivar ‘Fenditan’ were transformed through Agrobacterium tumefacien,expecting to get the transgenic chrysanthemum plants with improved resistance to waterlogging.After incubating,screening and differentiation,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 research on the effect of Vgb gene in improving chrysanthemum’s resistance to waterlogging.

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王 叶,高亦珂,张启翔,等.透明颤菌血红蛋白基因表达载体构建及转化地被菊研究[J].西北植物学报,2012,32(12):2390-2397

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  • 在线发布日期: 2013-01-06
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