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[Amplification of brain-derived neurotrophic factor receptor trkB gene and construction of its eukaryotic expression
Tao Huang1, Ru-xiang Xu, Lei Wu
1Department of Neurosurgery, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China. arteries@263.net
Objective:
To construct a recombinant eukaryotic expression vector of rat brain-derived neurotrophic factor receptor trkB gene.
Methods:
Using the total RNA extracted from rat brain tissue as the template, the trkB gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) with a pair of specific primers containing the restriction sites of EcoRI and BamHI. The amplified fragment of trkB gene was digested with EcoRI and BamHI, and then subcloned into cloning vector pMD18-T and then expression vector pEGFP-C2. The recombinant plasmid was identified by restriction endonuclease analysis and PCR.
Results:
The amplified DNA fragment was about 1 461 bp in length, and enzyme digestion and PCR analysis showed that trkB gene had been successfully cloned into the vectors pMD18-T and pEGFP-C2.
Conclusion:
The trkB gene of rat has been successfully amplified and cloned into the eukaryotic expression vector pEGFP-C2.
