Clone and expression of nuclear factor ?B p65 TAD
- VernacularTitle:核因子NF-?B p65亚基TAD的克隆及表达
- Author:
Chunli HOU
;
Dajun YING
;
Yong WEI
- Publication Type:Journal Article
- Keywords:
nuclear factor ?B;
p65 transcription activation domain;
clone;
expression
- From:Journal of Third Military Medical University
1984;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To acquire NF-?B p65 TAD(transcription activation domain) and construct its eukaryotic expression vector and express it in endothelial cells.Methods Human umbilical vein endothelial cells(HUVECs) were cultured and total RNA was extracted.The p65 TAD gene was amplified by RT-PCR.After sequencing,the p65 TAD gene was inserted into the eukaryotic expression vector pEGFP-N1 with the green fluorescence protein,named pEGFP-N1-p65 TAD.pEGFP-N1-p65 TAD was transfected into HUVECs and its expression was observed under fluorescence microscope and analyzed by Western blotting.Results p65 TAD(288-548) was cloned successfully.The constructed plasmid including p65 TAD gene was identical to the designed.p65 TAD gene was expressed successfully in HUVECs.Conclusion The construction of eukaryotic expression vector including p65 TAD gene and its expression in HUVECs are very successful.
