Construction of expression plasmid vectors containing the luciferase gene of an algae Dunaliella salina;

Highly efficient expression of luciferase gene cDNA eukaryotic expressing vector in Bacteria;

The eukaryotic expressing vector pGL 2 of luciferase gene cDNA can express efficiently in prokaryotic cells.

These DNA fragments were inserted into the pGL3-Basic which was the vector of luciferase reporter gene.

[Method] Clonging the promoters of CYP3A4 and CYP2B6 which contained the elements that hPXR, a kind of nuclear receptor, can recognize and bind, and inserting the trans-element to the upstream of firefly luciferase reporter gene of the pGL4.

Amplified apolipoprotein A-I(apoA-I) promoter gene(376 bp) containing essential transcription regulatory elements from human genomic DNA was fused upstream to the luciferase reporter gene and neomycin resistance gene in the constructed pGL3B-neo vector.

Methods The +495bp～+584bp of SCN5A promoter was amplified by polymerase chain reaction(PCR)and recombinated into pGL3-promoter luciferase report gene vector and transient transfected HEK293 cell and H9C2 cell.

coli chromosome,the luciferase report gene was knocked in lacZ site of chromosome lac operon by using Red recombination system and selection-counterselection kan/sacB technology.