Construction and expression of the eukaryotic expression plasmid of Human pEGFP/H2B
- VernacularTitle:人类pEGFP/H2B真核表达质粒的构建及表达
- Author:
Yuanjiu XU
- Publication Type:Journal Article
- Keywords:
H2B gene;
Green fluorescence protein;
HEK293 cell;
Chromosome segregation
- From:
Journal of Chongqing Medical University
2007;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct the eukaryotic expression plasmid of EGFP/H2B and express it in the Human embryo kidney cells(HEK293).Methods:Part H2B gene was obtained by RT-PCR from cultured HEK293 cell's whole RNA.The sequence encoding H2B was cloned into pEGFP-C1 to construct the eukaryotic expression vector.After confirmed by enzymatic digestion and sequencing,the recombination plasmid was transfected to HEK293 cells by lipofectamine technology for observation of the fusion protein's expression.Results:The recombination plasmid of pEGFP/H2B was constructed successfully.Compared with the dispersing green fluorescence among the whole cells in the control cells transfected with the mock plasmid(pEGFP-C1),the green fluorescence was concentrated in the nuclei of HEK293 cells transfected with pEGFP/H2B and distributed with the chromosomes in the mitotic cells.Conclusions:The successful construction of pEGFP/H2B recombination plasmids establishes an available survey system to study the chromosome segregation mechanism.
