豆球蛋白基因转化花叶芋的研究

Studies on the Transformation of Coladium bicolor with Legumin Gene

利用重组DNA技术,构建具有Ap^rCm^r的遗传标记并带有豆球蛋白的基因重组质粒,通过“三亲支配”,将豆球蛋白基因插入到植物基因工程载体pGV3850:1103neo DNA中。用改良的共培养法进行单子叶植物花叶芋的叶盘(叶柄)转化,在Km的选择压力下得到转化的再生植株。利用Southern吸印杂交,证明豆球蛋白基因整合到花叶芋的基因组中;通过胭脂碱分析,表明pGV3850:1103neo中的胭脂碱合成酶基因在再生植株中进行了表达。

By inserting the chloramphenicol resistance gene of pBR322 plasmid DNA into the HindⅢ-PstI sites of pUC18-leg plasmid DNA, we constructed the mediated plasmid vector pUB341 which contained the Ap^rCm^r genetic markers and legumin gene. The legumin gene then was introduced into the plant engineering vector pGV3850:1103neo by triparental matings and homologous sequence recombination. Using the modified leaf-disc co-cultivation method, the legumin gene was transferred to the stems of monocotyledon plant Caladium bicolor. Under the selection pressure of Km, transformed calli regenerated into whole plantlets. The results of nopaline derection and Southern blot hybridization showed that the legumin gene had been integrated into the genome of the transformed Caladium bicolor.