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棉花GDP-甘露糖-3',5'-异构酶基因的克隆及其表达分析 被引量:3
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作者 王江英 张宁 +1 位作者 司怀军 吴家和 《华北农学报》 CSCD 北大核心 2012年第1期12-17,共6页
根据棉花盐胁迫相关的EST序列设计特异性引物,利用3',5'-RACE技术,从棉花叶片中克隆出GDP-甘露糖-3',5'-异构酶(GDP-mannose-3',5'-epimerase,GME)基因,命名为GhGME。基因全长1 467 bp,开放阅读框1 131 bp,编码... 根据棉花盐胁迫相关的EST序列设计特异性引物,利用3',5'-RACE技术,从棉花叶片中克隆出GDP-甘露糖-3',5'-异构酶(GDP-mannose-3',5'-epimerase,GME)基因,命名为GhGME。基因全长1 467 bp,开放阅读框1 131 bp,编码376个氨基酸,分子量为42.39 6 kDa,等电点pI=6.291。利用荧光定量Real-time RT-PCR方法对棉花根、茎、叶和3种处理下该基因的表达特性进行分析,结果表明,GhGME在根、茎中高水平表达,在叶中表达量较低。在4℃低温和200 mmol/L NaCl处理下该基因表达上调,然而用100μmol/L GA3处理则表现为下调表达。GhGME的不同表达情况暗示该基因可能在棉花抗逆反应中起着重要的作用,为该基因在棉花中抗逆功能和应用研究提供了基础。 展开更多
关键词 棉花 基因克隆 GDP-甘露糖-3' 5'-异构酶 表达分析
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Establishment of the Agrobacterium-mediated Genetic Transformation System of Ginkgo biloba and the Construction of the Expression Vector of Gb-DXR
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作者 冯国庆 杨颖舫 +4 位作者 李郑娜 成瑜 杨春贤 陈敏 廖志华 《Agricultural Science & Technology》 CAS 2010年第3期28-32,114,共6页
[Objective] The research aimed to provide reference for increasing the genetic transformation efficiency of Ginkgo biloba mediated by Agrobacterium.[Method] Taking the mature embryos of Ginkgo biloba seeds as explants... [Objective] The research aimed to provide reference for increasing the genetic transformation efficiency of Ginkgo biloba mediated by Agrobacterium.[Method] Taking the mature embryos of Ginkgo biloba seeds as explants,after 48 hours' pre-cultivation on MS medium in the absence of phytohormone,GUS gene was transmitted into embryos of Ginkgo biloba mediated by three kinds of Agrobacterium.Transient expression of GUS gene activity was observed through histochemical staining,and the influencing factors of the expression of GUS gene were analyzed.And the expression vector of 1-deoxy-D-xylulose-5-phosphate reductoisomerase in the biosynthesis approach of biobalide precursor of Ginkgo biloba was constructed.[Result] A more suitable genetic transformation scheme was obtained as follows:taking embryos of Ginkgo biloba as explants,using EHA105 Agrobacterium with pCAMBIA1304+ for infection,co-culture for 3 days and GUS staining.The results showed that transient expression rate of GUS after transformation was higher.[Conclusion] The research provide a more effective method for further study on the transgene of Ginkgo biloba. 展开更多
关键词 Embryos of Ginkgo biloba AGROBACTERIUM-MEDIATED Genetic transformation GUS gene 1-deoxy-D-xylulose-5-phosphate reductoisomerase Expression vector
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