Construction of eukaryotic expression vector with brain-derived neurotrophic factor receptor trkB gene / 中华创伤杂志(英文版)

ABSTRACT

<p><b>OBJECTIVE</b>To construct an eukaryotic expression vector carrying rat brain-derived neurotrophic factor receptor trkB gene.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>The trkB gene of rat has been amplified and cloned into the eukaryotic expression vector pEGFP-C2.</p>