首页|农杆菌介导转化法构建转CTV-cp的枳壳植株

农杆菌介导转化法构建转CTV-cp的枳壳植株

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目的:采用农杆菌介导法,将能抑制病毒侵染的外壳蛋白基因转入枳壳,为通过基因转化进行枳壳的抗病育种提供依据和方法。方法:以枳壳实生苗上胚轴为材料,农杆菌菌株为EHA101,含有质粒载体pGA482GG。质粒上插入了35 s启动子驱动的外源目的基因——柑桔衰退病病毒外壳蛋白基因(CTV-cp)及GUS,NPTⅡ基因。结果:枳壳转化试验中,抗菌素采用头孢霉素较好,浓度为300 mg*L-1;以卡那霉素作为选择试剂,浓度为50 mg*L-1;转化获得的抗性植株,GUS检测结果,70.0%的植株呈阳性反应;Southern 杂交分析(Southern blot)鉴定证明外壳蛋白基因已整合到枳壳植株的核基因组中。结论:通过农杆菌介导的转化法,成功地获得了转外壳蛋白基因的枳壳植株。
Plant Regeneration from Agobacterium-mediated CTV-cp Gene Transformation of Poncirus trifoliata Raf.
Objective:To lay a foundation for the resistant breding, the anti-virus CTV-cp gene was transformed into the epicotyles mediated by Agrobacterium tumefaciens in Poncirus trifoliata. Method:The explants used for the genetic transformation were the epicotyls from P.trifoliata.The Agrobacterum tumefaciens strain was EHA101 containing vector plasmid pGA482GG.The coat protein gene (CTV-cp gene),GUS gene and NPT II gene were introduced into the transformation plasmid.Results:Ceftaxime used as antibiotics was better than carbenicillin. The concentration was 300 mg*L-1;The selection pressure for kanamycin was 50 mg*L-1;70.0% of the resistant plants were GUS-positive; extra gene was proved to be in P.trifoliata plant by southern blot examination. Conclusion:An effective genetic transformation mediated by Agrobacterium tumefacines,which harbours a CTV-cp gene, has been developed in P.trifoliata.Transgenic CTV-cp plants were obtained.

贺红、韩美丽、李耿光

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广州中医药大学 中药所

广西林业科学研究院

中国科学院 华南植物研究所

枳壳 外壳蛋白基因 植物转化

国家自然科学基金

39670476

2001

中国中药杂志
中国药学会

中国中药杂志

CSCD北大核心
影响因子:1.718
ISSN:1001-5302
年,卷(期):2001.26(1)
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