转Chi-Glu基因野罂粟植株卡那霉素筛选技术
Guizhou Agricultural Sciences(2010)
Abstract
为探索卡那霉素(Kan)抗性筛选在转基因野罂粟上的应用,以含有几丁质酶(Chi)和β-1,3-葡聚糖酶(Glu)的双价基因的T0代转基因野罂粟种子为试验材料,进行卡那霉素叶喷、叶片涂抹试验,筛选卡那霉素的最佳浓度;对T1代转基因植株进行卡那霉素筛选,并用PCR检验其阳性植株。结果表明:培养皿幼苗最佳叶喷浓度为50 mg/L;土培苗最佳叶喷和涂抹浓度为6.5g/L,连续叶喷或涂抹5d,每天1次。经卡那霉素筛选共获得12株转基因野罂粟植株,通过PCR检测,有4个株系已将Chi-Glu双价抗病基因整合进野罂粟的基因组中。
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Key words
screening,kanamycin,transgenosis,wild poppy,PCR
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