Carcinogenesis, Teratogenesis & Mutagenesis ›› 2004, Vol. 16 ›› Issue (1): 5-8.doi: 10.3969/j.issn.1004-616x.2004.01.002

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Constructing and Identifying the Firefly Luciferase Report Gene Vectors For 5' Flanking Regulated Elements of NGAL Gene

XU Li-yan, LI En-min, CAI Wei-jia , et al   

  1. Department of Oncologic Pathology,Medical College of Shantou University , Shantou 515031,China
  • Received:2003-10-07 Revised:2003-10-31 Online:2004-01-30 Published:2004-01-30
  • Contact: LI En-min

Abstract: BACKGROUND & AIM:To construct the firefly luciferase report gene vectorsfor5'flanking regulated elements of NGAL gene. MATERIAL AND METHODS:The DNA fragments G0~G6 (-1431~+84) of 5'flanking of NGAL gene were amplified from the genomic DNAs of esophageal cancer cell line SHEEC by using PCR. The PCR products were cloned into pGEM-Teasx vector , then directionally subcloned into pGL3-Basic vector. The recombined clones were identified by agarose gel electrophoresis after restriction endonuclease digesting. RESULTS:7 expression vectors , pGLB-G0~G6 , for 5'flanking regulated elements of NGAL gene had been constructed. CONCLUSION:These expression vectors offered the basic experimental conditions for studying the distribution characters of NGAL gene 5' flanking regulation elements and the properties of interactions between the elements and regulation proteins with the help of dual luciferase report gene assay system.

Key words: NGAL gene, regulation of gene expression, luciferase, report gene, construction vector