Li Xiangyun,Xu Xiang,Li Bingxu,Huang Hong,Xu Yunsheng.Construction of human eIF4A3 eukaryotic expression plasmid and screening of stable cell line[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(18):1897-1899.
Li Xiangyun,Xu Xiang,Li Bingxu,Huang Hong,Xu Yunsheng.Construction of human eIF4A3 eukaryotic expression plasmid and screening of stable cell line[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(18):1897-1899.DOI:
Construction of human eIF4A3 eukaryotic expression plasmid and screening of stable cell line
Objective To construct the human eIF4A3 recombinant vector and screen the stable cell line over-expressing human eIF4A3. Methods Human eIF4A3 gene was cloned by RT-PCR. Gene fragments were linked to HA and c-myc labeled plasmid vectors to transfect HeLa cells. A stable cell line was generated by screening G418. In vitro translation was conducted in an in vitro translation system. Expression of eIF4A3 was identified by Western blotting. Results The human eIF4A3 eukaryotic expression vector was successfully constructed
which was stably expressed in HeLa cells and could translate the eIF4A3 protein in an in vitro translation system. Preliminary function study showed that eIF4A3 could inhibit the expression of luciferase reporter gene. Conclusion A HeLa cell line over-expressing human eIF4A3 we generated can be used as an eukaryotic expression vector in an in vitro translation system.