Cloning and construction of eukaryotic expression vector for rat neurotrophin 3 gene

Objective To clone and construct the eukaryotic expression vector for rat Neurotrophin 3(NT-3)gene.Methods The rat NT-3 cDNA was amplified by RT-PCR from rat brain tissue.By gene recombination technique,rat NT-3 cDNA was inserted into eukaryotic expression vector pcDNA3 to construct recombinant plasmid pcDNA3-N3.The recombinant plasmid was identified with restriction enzyme digestion and DNA sequencing.Results The RT-PCR product is 822bp specific segment.By restriction enzyme digestion,the recombinant plasmid was digested into 822bp and 5200bp fragments.The DNA sequence of the 822bp fragment was identical with rat NT-3 cDNA in GenBank.Conclusion The NT-3 gene was cloned,and the eukaryotic expression vector for NT-3 gene was constructed successfully,which will provide the foundation for the further research.