[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 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.
Key Laboratory of Molecular Biophysics of the Ministry of Education, China-UK Joint Lab, College of life Science & Technology, Huazhong University of Science & Technology Wuhan, 430074, China.
Zhi-Ming
ZM
Tu
Lin
L
Chen
Guang-Xiao
GX
Yang
Guang-Yuan
GY
He
D000469:Alkaline Phosphatase / Q000378:metabolism*; D015342:DNA Probes / Q000235:genetics / Q000378:metabolism*; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics; D009693:Nucleic Acid Hybridization; D010761:Phosphorus Radioisotopes / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D019076:Transgenes / Q000235:genetics
D015342:DNA Probes; D004279:DNA, Viral; D010761:Phosphorus Radioisotopes; D000469:Alkaline Phosphatase