Construction and identification of the lentivirus vector expressing IL- 1ra

Objective To construct the lentivirus vector expressing IL - 1 ra,which will be used in the study of gene modification and transplantation of neural precursor cells (NPCs).Methods IL -1ra cDNA was cloned from IL - 1ra plasmid by PCR,fused with IRES2 - EGFP downstream,and loaded to pLenti6.3 V5 - DEST.After sequencing and package were completed,the lentivirus plasmid was named as pLenti6.3-IL1ra -IRES2 - EGFP and was used to infect HEK293 cells.The titer of virus was tested according to the expressing level of EGFP.Results DNA sequencing analysis confirmed that the DNA sequence of IL -lra in plasmid amplified from primers CMV - F and IRES2 - R was the same as which was adopted in Genebank.The titer of the virus suspension was 3.5 × 106 TU/ml.There was not obvious change of growth characteristics in NPCs after which were transinfected with recombinant lentivirus.Conclusion Lentivirus vector expressing IL -1ra was constructed successfully. Key words: IL- 1ra;  Lentivirus vector;  Gene modification;  Cell transplantation;  Spinal cord injury