Effect Of Recombinant Pyrococcus Yayanosii CH1-ferritin On Iron Supplementation In Female SD Rats With Iron Deficiency Anemia

Ferritin is an important biological metabolic protein in iron storage process, it has the function of iron storage and iron removal. Ferritin can be found in most of the organisms and it also can tolerate the dilute acid pH=2.1, dilute alkali pH=0.1 and high temperature 70～75℃. Ferritin is composed of an outer protein shell and an inner iron core.The ferritin shell contains 24 subunits that forms the hollow spherical structure. The diameter of ferritin shell is 12 nm and the diameter of iron core is 8 nm. Iron deficiency anemia (IDA) is caused by iron deficiency and lead to a low hemoglobin level.Objective:This study constructed a high expression strain of BL21-CHI-ferritin by gene engineering technology that could express recombinant CH1-ferritin efficiently. And the study also took preliminary exploration on the physical and chemical properties of recombinant CH1-ferritin. Finally, iron supplementation trials were carried out on rat model with IDA by gastric perfusing natural mineralized recombinant CH1-ferritin. The purpose of gastric perfusion trials were to testify the effect of recombinant CHI-ferritin on iron supplementation and provide references for the future clinical research.Methods:Genome of Pyrococcus yayanosii CH1 and the gene sequence of CH1-ferritin were found and located in GenBank. Then the CHI-ferritin codon preference was modified and the CH1-ferritin gene sequence was synthesized by using gene synthesis method. Next we constructed a high expression strain of BL21-CH1-ferritin that could express recombinant CH1-ferritin efficiently. We used natural mineralization method to make the recombinant CH1-ferritin form an inner iron core. Finally we constructed an iron deficiency anemia female SD rat model and the model was gastric perfused recombinant CH1-ferritin to verify the effect of recombinant CH1-ferritin on iron supplementation.Results:We successfully constructed the efficient expression strain of BL21-CH1-ferritin and purified the recombinant CH1-ferritin by cntroling temprature. And we got the result of preliminary exploration on the physical and chemical properties of recombinant CH1-ferritin. Finally, we used natural mineralization method to make the ferritin form an inner iron core. Then gastric perfusion trail on the rat model with IDA indicated that the natural mineralized CH1-ferritin could pose a good effect on iron supplementation and CH1-apoferritin couldn’t affect iron supplementation.Conclusion:Synthetic gene sequence of CH1-ferritin dosen’t change amino acid sequence and it has the same ability to express biological ferritin. Naturally mineralized CH1-ferritin took a good effect on iron supplementation.