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HEREDITAS ›› 2007, Vol. 29 ›› Issue (12): 1533-1537.doi: 10.1360/yc-007-1533

• 技术与方法 • Previous Articles    

Application of enzyme-labeled probe in testing of transgenic plant

TU Zhi-Ming; CHEN Lin; YANG Guang-Xiao; HE Guang-Yuan

  

  1. 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
  • Received:2007-04-26 Revised:2007-07-08 Online:2007-12-10 Published:2007-12-10
  • Contact: HE Guang-yuan

Abstract:

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.