Cloning and Identification of 5'Flanking Region of Promoter in NGAL Gene

Objective 　Cloning and identifing of 5'flanking region promoter in NGAL ( neut rophil gelatinase - associated lipocalin) gene. Methods 　Two different length f ragments -416～+ 84 and -152～+84 of 5'flanking region of NGAL gene were cloned into pGL3-Enhancer vector (pGLE), which aids in verification of functional promoter element s. And pGLE-416 and pGLE-152 reporter gene vectors were const ructed. Then they were respectively cot ransfected with pRL-TK vector into HeLa, EC109 and Vero cells. Relative light unit (RLU) in the cells was measured with Dual Luciferase Report Gene Assay System (DLR) to prove whether there are promoter element s in these f ragments. Results 　RLU of pGLE-152 was obviously increased (P<0. 05), compared with control pGLE, after they had been transfected into three different cell lines. Moreover, there was variable fold induction in the different cells. Conclusion 　The promoter of NGAL gene is located in-152～+84 region and has a cellular specialty, which might relate to cooperation with enhancer elements.