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Published on: November 1, 2014
[Cloning and expression of the gene encoding rat IgE-dependent histamine-releasing factor and effect study on the
Xiao-xiang Chen1, Xu-chu Hu, Jin Xu
1Department of Parasitology, Preclinical School of Sun Yat-sen University, Guangzhou 510080, China. chenxiaoxiang75@yahoo.com.cn
Aim:
To clone and express the gene encoding rat IgE-dependent histamine-releasing factor (rHRF) and to study the effect of recombinant rHRF to induce histamine release from rat sensitized mast cells.
Methods:
The complete coding region of rHRF was cloned and expressed in BL21 cells. The soluble recombinant rHRF was purified. Aliquots of the mast cells obtained from the lungs of OVA-immunized rats were separately stimulated by recombinant rHRF at different concentration. The released histamine was measured by the OPT spectrofluorometric procedure.
Results:
The cloned DNA fragment showed 97% nucleotide sequence identity and 95% deduced amino acid sequence identity with the mRNA of rat IgE-dependent histamine-releasing factor (or translationally controlled tumor protein) in GenBank. The recombinant rHRF showed a relative molecular mass of 24 000 in SDS-PAGE. The purified rHRF which was independent of the allergen induced histamine releasing from the sensitized mast cells in a dose-dependent manner.
Conclusion:
The recombinant rHRF, which showed the effect of inducing histamine release from sensitized mast cells, would play an important role in the allergen diseases.
