Cloning and Prokaryotic Expression of NtCCHC in Narcissus tazetta var. chinensis

  Objective  Based on the information in the SSH Library on Narcissus tazetta var.chinensis, the zinc finger gene was cloned and prokaryotic expressed for future studies.

  Method  Total RNA of N.tazetta var. chinensis from the paclobutrazol-treated leaves was extracted using the Trizol method. After reverse transcription into cDNA, gene cloning and homology analysis were carried out on it with primers designed according to the known sequence of CCHC. A prokaryotic expression vector was constructed to induce expression.

  Result  The 810 bp sequence encoding 269 amino acids was cloned. The cloned gene showed high homologies with the CCHC genes of many species and was named NtCCHC, which could be successfully induced and expressed in pGEX-4T-3. The phylogenetic tree constructed by MEGA 6.0 indicated that the closest genetic distance of NtCCHC lied with Nelunbo nucifera.

  Conclusion  This study successfully cloned and expressed the NtCCHC gene sequence of N. chinensis.