Promotion effiency of EGFP is influenced by mutation of hTERTp

Objective To observe the fluores cence in tensity change of enhanced green fluo rescent protein(EG FP)promoted by mutated human telomerase reverse transcriptase(hTERT)promoter.Methods The hTERT mutated promoter with 300 bp were synthesized with PCR method including two single E-box mutation sequence,a double E-box mutation sequence,five single Sp1 binding site mutation sequence,and a whole Sp1 binding site mutation sequence.The ten hTERT promoters sequence were all inserted into free-promoter EGFP upstream to reconstruct a recombinant plasmid.And the fluorescent in tensity was detected.Results All mutated hTERT promoters could induce the decrease of fluore scent intensity except one-110bp single Sp1 mutation promoter.The fluorescent intensity of double E-box mutation promoter and five Sp1 bingd in site mutation promoter were decreased obviously than the control.Conclusion The EGFP fluore scent intensity were decreased after transfected mutated hTERTp gene in SW 579 cells,which proved the E-box and SP1 binding site sequence of hTERT pare correlated with the promotion efficiency.