HNGFR (TrkA) gene cDNA cloning by pGEM -Easy T-Vector and Sequencing

Objective:To construct the pGEM-Easy T-vector containing HNGFR (NGF High affinity Receptor,TrkA) gene.Methods:By following total RNA being extracted from dorsal root ganglia (DRG) of rat and RT-PCR being carried out, pGEM-Easy T-vector was linked with PCR products. After E. coli DH5α was transformed with recombinants and screened with blue/ white blot test for positive clones from which plasmids were then extracted, the recombinants were identified by restricted analysis and sequencing. Results: 416 base pairs fragment was obtained through RT-PCR. After cloning by pGEM-Easy T-vector and being identified by restricted analysis and sequencing, it is confirmed that the object fragment has 100% homology with TrkA sequence in Genbank. Conclusion:The T-vector clone with the conservative region of TrkA gene has been constructed successfully. This clone can be used in neuronal toxicological research area such as some metals poisoning.