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利用发根农杆菌(Agrobacteriom rhizogenes)介导二元载体向番茄导入TMV,CMV外壳蛋白基因的研究 被引量:14

STUDIES ON TMV CP AND CMV CP GENES CARRIED INTO TOMATO BY BINARY VECTOR WITH AGROBACTERIUM RHIZOGENES
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摘要 本文用含PRi(PRiA4b)的发根农扦菌为介导,将二元载体质粒PBTC-8(T—DNA上有TMV—CP和CMV—CP基因),导入黑龙江省当地番茄品种。用胚轴注射,顶端切口涂抹,子叶切块穿刺等方法感染转化子菌液,诱导出毛状根。滤纸电泳检测有50%以上的毛状根含有农抒诚和甘露碱。用抗性筛选法亦获得有卡那霉素抗性的毛状根。在穿刺感染子叶切块诱导产生的具卡那霉素抗性的愈伤组织上,获得不定芽与再生植株,经冠瘿碱,PCR,斑点免疫结合法检测证明再生植株中有TMV和CMV基因产物的两种外壳蛋白和农扦碱与甘露碱,获得双转化植株。 Using Agrobacterium rhizogenes with Ri-plsmid (PRiA4b) to be medium,the binary vector PBTC 8 (T-DNA with CMV-cp and TMV-cp genes) was transforred in to cultivated tomato of Heilongiiang province. The method of hypocotyl,apical half of epicotyl and cutting cotyledon sections was used to induct hairy roots. Agropion and mannopine were detected in half of these hairy roots by electrophores. Some hairy roots were get by the method of sdlection medium with kanomycine resistance. The adventitious bud and regeneration plant were induced from cullus with kanomycine resistance on infected cotyledon piece. Detecting the alkaloid, PCR and DIBA (Dot Immunobinding Assy),it certified there were TMV-cp and CMV-cp genes, Agropion and mannopin in the regeneration plant We got deuble traslative plant.
机构地区 哈尔滨师范大学
出处 《植物研究》 CSCD 北大核心 1994年第4期416-423,共8页 Bulletin of Botanical Research
关键词 番茄 CMV-CP基因 发根农杆菌 载体 Ri plasmid Tomato TMV-cp and CMV-cp genes.
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