[Expression of fused luxAB gene of bacterial luciferase in liver carcinoma cells].
OBJECTIVE
This work was done to look into the expression of fused luxAB gene of bacterial luciferase as a reporter gene in liver carcinoma cells.


METHODS
The mammalian expression vector pcDNA3-luxAB gene, constructed by the fusion of luxA and luxB genes, were amplified in the polymerase chain reaction (PCR) directed site mutagenesis from the vibrio harveyi 1600 strain and inserted into the plasmid of pcDNA3. This analysis was to confirm the fused luxAB gene and the positive clones obtained by the G418 resistant stable selection and transfected by lipofectin, when they were confirmed by the PCR. The growth curve of cell population and luminescence of bacterial luciferase was obtained through MTT and bioluminescence, respectively.


RESULTS
The fused luxAB gene, being a monocistron, completely agreed with the design. No significant difference in the growth curves of cell population was observed between the transfected cells and untransfected ones. The recombinant plasmid was likely to be expressed in a stable fashion in the BEL7402 cell. Meanwhile, the maximum cellular level in terms of vitro bioluminescent strength reached the point of (8.71 +/- 1.21) mV/40 micrograms protein.


CONCLUSIONS
Bacterial luciferase luxAB gene may become the first choice as a new, sensitive and non-invasive reporter gene in the research on liver cancer cells.
Mao
C
C
Beijing Hospital, Beijing 100730, China.
Yang
S
S
Zhao
M
M
D000818:Animals; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D002455:Cell Division; D002999:Clone Cells; D004247:DNA; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D008322:Mammals; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection; D014407:Tumor Cells, Cultured
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D004247:DNA; D008156:Luciferases