Construction the eukaryotic expression vector of human decay accelerating factor and transfection NIH/3T3 cells
- VernacularTitle:构建人补体衰变加速因子真核载体并转染NIH/3T3成纤维细胞
- Author:
Qing QIAO
;
Yong CHEN
;
Kefeng DOU
;
Jing ZHANG
;
Jianpin LI
- Publication Type:Journal Article
- Keywords:
Decay accelerating factor;
Chitosan;
Nanoparticle;
Gene clone;
Gene transfection
- From:
Journal of Medical Postgraduates
2003;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct eukaryotic expression vector pSecTag2/HygroB-DAF of human decay accelerating factor (DAF) and transfect NIH/3T3 cells after encapsulated by chitosan. Methods:The human DAF fragments were obtained by PCR form DAF-pGEM-T Easy Vector, cloned into the eukaryotic expression vector pSecTag2/HygroB, and identified by restriction endonuclease’s digestion and DNA sequencing. After the particles of pSecTag2/HygroB-DAF were encapsulated by chitosan, the NIH/3T3 cells were transfected by chitosan-DAF nanoparticles and detected DAF expression by immunohistochemistry stain. Results:The DAF fragment was 1049 bp. Its sequence was as same as DAF cDNA in Genebank. After having been transfected by chitosan-DAF nanoparticles 24 hours, the NIH/3T3 cells showed diffusely positive in cytoplasms by anti-DAF immunohistochemistry. Conclusion:Eukaryotic expression vector of human DAF were constructed successfully and transfected it to NIH/3T3 cells after encapsulated by chitosan.
