Construction and identification of recombinant TPH2-targeted small interfering RNA expression vector

Objective To construct and identify recombinant TPH-2-targeted small interfering RNA (siRNA) expression vector.Methods Oligonucleotide containing the small hairpin of TPH-2 was designed and synthesized,which was inserted into the pU6-CMV-GFP plasmid double digested by Age Ⅰ and EcoR Ⅰ.The aim plasmids were transformed into competent cells E.coli DH5α.The grown colonies were identified by colony polymerase chain reaction (PCR) and then the positive colonies were sequenced and aligned.Results The result of PCR and gene sequencing confirmed that the pU6-CMV-GFP-TPH-2 shRNA expression vector with 332 bp.Conclusion The expertment had been constructed successfully. Key words: Tryptophan hydroxylase;  RNA interfere;  Expression vector