酶标探针在转基因植物检测中的应用

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。

Application of enzyme-labeled probe in testing of transgenic plant

Used alkaline-phosphatase-labeled DNA as a probe to examine the expression of foreign UidA gene in trans-genic 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.