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Published on: December 22, 2014
Construction of eukaryotic expression vector with brain-derived neurotrophic factor receptor trkB gene
Tao Huang1, Xiao-dan Jiang, Zhong Xu
1Department of Neurosurgery, Zhujiang Hospital, Nanfang Medical University, Guangzhou 510282, China.
Objective:
To construct an eukaryotic expression vector carrying rat brain-derived neurotrophic factor receptor trkB gene.
Methods:
Using the total RNA isolated from rat brain as template, the trkB gene was amplified by reverse-transcription-polymerase chain reaction (RT-PCR) with a pair of specific primers which contained the restrictive sites of EcoR I and BamH I. The amplified fragment of trkB gene was digested with EcoR I and BamH I, and then subcloned into cloning vector pMD18-T and expression vector pEGFP-C2 respectively. The recombinant plasmids were identified by restriction endonuclease enzyme analysis and PCR.
Results:
The amplified DNA fragment was about 1461 bp in length. Enzyme digestion and PCR analysis showed that the gene of trkB had been successfully cloned into vector pMD18-T and pEGFP-C2.
Conclusions:
The trkB gene of rat has been amplified and cloned into the eukaryotic expression vector pEGFP-C2.
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