酶标探针在转基因植物检测中的应用  

Application of enzyme-labeled probe in testing of transgenic plant

作　　者：涂知明[1] 陈泠[1] 杨广笑[1] 何光源[1] 

机构地区：[1]华中科技大学生命科学与技术学院中英联合实验室分子生物物理学教育部重点实验室,武汉430074

出　　处：《遗传》2007年第12期1533-1537,共5页Hereditas（Beijing)

基　　金：国家重点基础研究发展计划(973计划)(编号:2002CB111302);国家自然科学基金(编号:30370807);华中科技大学博士论文基金资助项目~~

摘　　要：采用碱性磷酸酶标记DNA制备分子探针,并首次在植物中应用。酶在苯醌作用下与单链DNA联结,形成DNA和酶的共价复合物即酶标探针。此探针通过分子杂交与待测DNA结合,再与酶的底物作用显色,3～6h内可观察结果。用此探针检测转基因植物中的UidA基因,点杂交和Southern杂交结果表明,所合成的酶标探针具有快速、准确、安全而经济的优点。点杂交证明外源UidA基因被成功转化到受体植物中,Southern杂交对转基因的材料检测的结果证明,该材料包含多个外源UidA基因拷贝,初步确定其外源UidA基因拷贝数在5个以上。Used alkaline-phosphatase-labeled DNA as a probe to examine the expression of foreign UidA gene in transgenic plants. Alkaline phosphatase coupled with polyethyleneimine （PEI） using P-benzoquine as the cross-linking reagent was covalently linked to single-stranded DNA via glutraldehyde. Such DNA-enzyme complexes were used as a probe for dot hybridization and Southern blot. After hybridization and incubation with a substrate solution, results can be observed directly in three to six hours and the results showed that it was a sensitive, specific, rapid, safe and economical probe. Dot hybridization analysis showed that the UidA gene was transformed into the target plants and southern blot showed that there were at least 5 copies of UidA gene in transgenic plants.

关 键 词：组织特异性启动子 洲烈基因 杂交 酶标探针 

分 类 号：Q943.2[生物学—植物学]