酶液处理法制备转基因植株PCR DNA模板的研究  

Study on enzyme-treated preparation of DNA template for PCR assay of transgenic plants

作　　者：郎春秀[1] 陈笑芸[1] 王伏林[1] 吴关庭[1] 胡张华[1] 陈锦清[1] 

机构地区：[1]浙江省农业科学院病毒学与生物技术研究所,浙江杭州310021

出　　处：《浙江农业学报》2005年第5期337-340,共4页Acta Agriculturae Zhejiangensis

基　　金：国家自然科学基金项目(30370910);浙江省自然科学基金项目(M303144)

摘　　要：采用脂肪酶、蛋白酶K和复合酶处理叶片,建立了酶液处理法制备转基因植株PCR DNA模板的新技术并在转基因油菜和水稻上进行了验证.酶液浓度为0.1 mg/ml脂肪酶、0.1 mg/ml蛋白酶K和0.1 mg/ml复合酶,叶片量为1～2 cm2时,所获得的模板质量最高,PCR扩增效果最佳.该方法操作简便、稳定性较高、结果准确可靠,适用于大批量转化植株和转基因后代植株的PCR DNA模板制备.A new method for preparation of DNA template for PCR assay of transgenic plants was established by treating leaves with lipase, protease K and complex enzyme and was demonstrated in transgenic rape and rice. The optimun concentration of three enzymes was 0.1 mg/ml and the appropriate leaf area was 1 - 2 cm^2. With advantages of simplicity in operation, high stability and high reliability, the new method could be used for DNA template preparation of a large quantity of transformed or transgenic progeny plants.

关 键 词：酶处理 DNA模板 制备 转基因植株 PCR检测 

分 类 号：S562[农业科学—作物学]