Cloning of the Murine Na+-K+-2Cl-Cotransporter Gene Promoter and the Effect of 20-HETE on Its Transcriptional Activity / 中国医科大学学报
Journal of China Medical University
;
(12): 29-33, 2019.
Artigo
em Chinês
| WPRIM
| ID: wpr-744793
ABSTRACT
Objective To clone the murine Na+-K+-2Cl-cotransporter (Nkcc2) gene promoter and analyze 20-HETE regulation of the murine Nkcc2 gene transcriptional activity. Methods A fragment of the murine Nkcc2 gene promoter was analyzed using bioinformatics software AliBaba and TRANSFAC TESS. The murine Nkcc2 gene promoter fragment (-1 462 bp-+40 bp) was amplified by PCR using murine genomic DNA as a template and then cloned into a pGL3-Basic vector to generate a luciferase reporter construct. The recombinant reporter construct was transiently transfected into HEK293 T cells using Lipofectamine 2000 for 24 h. The transfected HEK293 T cells were treated with 20-HETE for 2 h followed by measurement of the luciferase activity using the Dual-Luciferase Reporter Assay system. Results A luciferase reporter construct containing the murine Nkcc2 gene promoter was successfully generated. The results showed that 20-HETE significantly reduced the transcriptional activity of the construct. Conclusion 20-HETE may reduce the expression of the murine Nkcc2 gene through transcriptional regulation.
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Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Journal of China Medical University
Ano de publicação:
2019
Tipo de documento:
Artigo
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