Transient expression assay of [A]gamma-588 [A/G] mutations in the k562 cell line
IBJ-Iranian Biomedical Journal. 2011; 15 (1,2): 15-21
in English
| IMEMR
| ID: emr-129772
ABSTRACT
In the previous study, we have shown that the presence of A allele at position -588 in [A]gamma -globin gene was highly frequent and closely associated with fetal hemoglobin elevation among beta-thalassemia intermedia patients. Therefore, we decided to investigate whether this allele [A allele at -588] could result in an increase in [A]gamma-globin gene expression to ameliorate the severity of the disease in thalassemia patients. Three constructs containing ji locus control region, [A]gamma -globin and beta-globin genes were designed and employed in the transient expression assay. The difference among constructs was in the promoter region of, [A]gamma -globin gene [A and G alleles at -588]. A construct with T to C base substitution at -175 of, [A]gamma -globin, created by site-directed mutagenesis, was selected as positive control. The K562 cell line was transfected with the above constructs. Subsequently, the expression of, [A]gamma -globin gene was determined by quantitative real-time reverse transcription-PCR. There was not a significant increase in the expression of, [A]gamma -globin gene in the construct containing A allele comparing the one with G allele at -588. -588 [A>G] mutation does not play a major role in regulation of, [A]gamma -globin gene, suggesting that other factors may be involved
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Index:
IMEMR (Eastern Mediterranean)
Main subject:
Gamma-Globulins
/
RNA, Messenger
/
Transfection
/
Gene Expression Regulation, Leukemic
/
Genetic Techniques
/
K562 Cells
/
Flow Cytometry
/
Mutation
Limits:
Humans
Language:
English
Journal:
Iran. Biomed. J.
Year:
2011
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