Construction and identification of recombinant firefly luciferase reporter plasmid pGL2-PEPCK-Luc.
- Author:
Kai FENG
1
;
Heng WANG
;
Qi SUN
;
Xin-hua XIAO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Base Sequence; Gene Expression Regulation, Enzymologic; drug effects; Humans; Liver Neoplasms; genetics; Luciferases; genetics; Molecular Sequence Data; Phosphoenolpyruvate Carboxykinase (GTP); genetics; metabolism; Plasmids; genetics; Promoter Regions, Genetic; genetics; Protein Binding; Rats; Recombinant Fusion Proteins; genetics; metabolism; Transcription, Genetic; Transfection; Tumor Cells, Cultured
- From: Acta Academiae Medicinae Sinicae 2004;26(5):562-565
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo evaluate in vitro regulation of phosphoenolpyruvate carboxykinase (PEPCK) gene promoter on gene transcription, and construct luciferase reporter plasmid pGL2-PEPCK-Luc.
METHODSA 550 bp fragment of PEPCK promoter cut from plasmid pPEPCK-int was inserted into transitional vector PBS-SK to construct a transition plasmid PBS-PEPCK. Then the recombinant luciferase reporter plasmid pGL2-PEPCK-Luc was cloned.
RESULTSRestriction enzymes and nucleotide sequence conformed that the coupling site of recombinant plasmid was correct without base mutation and deletion, and the sequence inserted was the same as data of GeneBank. The luciferase could be expressed in hepatoma cell transfected by pGL2-PEPCK-Luc.
CONCLUSIONEstablished a new means to study transcriptional regulation of PEPCK promoter.
