[Transient and stable expression of human ApoE7 gene in NIH/3T3 cell lines].
OBJECTIVE
To investigate the expressing profile of h-apoE7 gene under the regulation of MT-I promoter in mouse cells.


METHODS
The eukaryotic expressing plasmid pME7 was constructed through cloning human apoE7 genomic DNA(6.0 kb) into pMT at the site of Bgl II/Hind III just under the control of the mouse metallothionein-I promoter. Transfection of pME7 plasmid into NIH/3T3 cell lines by Lipo-fectamine was performed.


RESULTS
The transient expression of recombinant pME7 in NIH/3T3 cells showed that the expression was not restricted to human cells; The stable expression of pME7 demonstrated that the levels of h-apoE7 mRNA was highly related to the copies of integrated h-apoE7 gene. Whereas the results of ELISA implicated that the exogenous human apoE7 gene was transcribed, processed properly, and eventually translated into functional proteins. Furthermore, heavy metal (ZnSO4) could enhance the expressing level of h-apoE7 gene in transfected cell lines. The level of human apoE7 protein arised 45%-60% while induced.


CONCLUSIONS
It was suggested that MT-I promoter could modulate the expression of recombinant h-apoE7 gene correctly. Therefore, cellular studies provided the scientific basis for establishment of h-apoE7 transgenic mouse models.
Sun
M
M
Institute of Basic Medical Sciences, National Laboratory of Medical Molecular Biology, CAMS and PUMC, Beijing 100005.
Qi
Z
Z
Pan
Y
Y
D016475:3T3 Cells / Q000378:metabolism; D000818:Animals; D053318:Apolipoprotein E3; D001057:Apolipoproteins E / Q000096:biosynthesis / Q000235:genetics*; D016678:Genome; D006801:Humans; D008668:Metallothionein / Q000235:genetics; D051379:Mice; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014162:Transfection
D053318:Apolipoprotein E3; D001057:Apolipoproteins E; C049579:apolipoprotein E7, human; D008668:Metallothionein