Cloning of human ? thalassemic mutation ? IVS II654(C→T) and establishment of its mammalian expression system / 中国病理生理杂志
Chinese Journal of Pathophysiology
;
(12)1986.
Article
in Chinese
| WPRIM
| ID: wpr-520269
ABSTRACT
AIM:
To clone human ?-globin gene carrying a thalassemic mutation IVS II654(C→T) and establish a eukaryotic expression system for high-level expression of human ? IVS II654 gene in mouse erythroleukaemia(MEL) cells.METHODS:
The fragments of human ? 654 gene isolated from the ? thalassemia patients homozygous for the ? 654 mutation were amplified by PCR, and cloned to plasmid pBGT51. Then, the human ? LCR and ? 654 gene were subcloned from plasmid pBGT51 to the stable mammalian expression vector pcDNA3.1+ together, and the MEL cells were transfected with this vector using commercially available cationic lipid FuGENE6. The MEL cells were induced for further maturation by DMSO and the expression of human ? 654 gene in the MEL cells was identified by RT-PCR.RESULTS:
A mammalian expression system of human ? thalassemic mutation ?IVS II654(C→T) was established.CONCLUSION:
The level and the reliability of expression of human ? 654 gene in the MEL cells in vitro are similar to that in vivo in human body. This may be a valuable gene therapy model for human ? thalassemic mutation ?IVS II654(C→T).
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Pathophysiology
Year:
1986
Type:
Article
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