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中国农学通报

中国农学通报 ›› 2014, Vol. 30 ›› Issue (6): 124-128.doi: 10.11924/j.issn.1000-6850.2013-1485

所属专题： 油料作物

• 农学 农业基础科学 • 上一篇    下一篇

花生抗黄曲霉的植物双价表达载体的构建

肖宇 陈湘瑜 陈华 陈剑洪 庄伟建   

  • 收稿日期:2013-05-27 修回日期:2013-11-23 出版日期:2014-02-25 发布日期:2014-02-25
  • 基金资助:
    科技部国际科技合作计划项目“花生抗黄曲霉基因工程及育种研究”(2008DFA31450)；国家863计划项目“花生抗病功能基因组研究”(2013AA102602-5)；福建省科技厅高校产学合作科技重大项目“花生优异种质创制和高优新品种选育与产业化示范”(2011N51010064)；福建省科技厅项目“福建省作物分子与细胞生物学重点实验室”(2008I0102)。

Construction of Plant Bivalent Expression Vectors Against Aflatoxin in Peanut

  • Received:2013-05-27 Revised:2013-11-23 Online:2014-02-25 Published:2014-02-25

PDF (PC)

625

摘要： 通过常规育种培育的抗黄曲霉品种皆表现抗性不稳定，鉴此开展通过基因工程手段以培育高抗以及无筛选标记的转基因花生品种。几丁质酶基因CHI和葡聚糖酶基因GLU皆为广谱性的抗病基因且具有协同增效作用。分别以pSC1300-8-CHI和pSC1300-13-GLU载体为基础，通过PCR技术在CHI基因的特异启动子8#前端和T-nos末端分别加入NcoⅠ酶切位点和AflⅡ酶切位点，在GLU基因的特异启动子13#前端和T-nos末端分别加入ApaⅠ酶切位点和SpeⅠ酶切位点，通过酶切、连接将上述2个基因连接在抗生素自我删除载体pLoxp上，获得了具有抗生素自我删除选择标记的双价果种皮特异表达载体pLoxp-8-CHI-13-GLU。经过限制性内切酶鉴定，该植物表达载体构建成功。

关键词: 河北省, 河北省

Abstract: The performance of species resistance to Aspergillus by conventional breeding are unstable, therefore, genetic engineering is used to breed varieties resistant to Aspergillus. CHI and GLU are broad-spectrum resistance genes and have synergistic effect. Based on constructed vectors pSC1300-8-CHI and pSC1300-13-GLU, NcoⅠrestriction site was added to the front of the promotor 8# of CHI gene and AflⅡrestriction site was added to the end of T-nos sequence through PCR amplification, ApaⅠrestriction site was added to the front of the promotor 8# of GLU gene and SpeⅠrestriction site was added to the end of T-nos sequence through PCR amplification, two genes will be connected to antibiotics self-removing carrier pLoxp after digestion, antibiotic induced-removing plant bivalent expression vectors pLoxp-8-CHI-13-GLU was constructed. Identified by restriction endonuclease, the expression vector was successfully constructed.