A study on cloning, expressing and investigating biological activity of recombinant chicken interferon produced from yeast Pichia pastoris

Vo Thi Minh Tam, Nguyen Thi Thanh Giang, Nguyen Dang Quan, Nguyen Quoc Binh

Abstract


Virus-infectious diseases are permanent threats and cause serious lost for chicken industry of our country. Thus, the development of a bio-product enhancing the immune system against viral-caused diseases on chicken is highly necessary. Aims of this study were to clone, to express and to investigate anti-viral activity of recombinant chicken interferon-α and interferon-γ (ChIFN) produced from yeast Pichia pastoris. Gene fragments encoding ChIFN-α and ChIFN-γ were obtained by PCR and PCR/SOE (Splicing Overlapping Extension) based on the chicken genome DNA. Expressing vectors pPIC9K bearing gene-encoding ChINF-α or ChIFN-γ were cloned and two clones of P. pastoris stably expressing ChIFN-α and ChIFN-γ were generated.  The expression of ChIFN-α and ChIFN-γ in P. pastoris culture supernatant was identified by Tricine SDS-PAGE. The biological activity of recombinant ChIFN-α was investigated on the model of chicken embryo fibroblast infected with IBDV and NDV. In case of IBDV-infected cells, the survival rate of cells treated by 1.6 µg/ml ChIFN-α was 79-88%, significantly higher than that of cells treated by PBS which was 33-34%. With NDV-infected cells, the survival rate of cells treated by 1.6 µg/ml ChIFN-α was 81-90%, also obviously higher than that of cells treated by PBS which was 27-32%. In conclusion, recombinant ChIFN-α and ChIFN-γ were expressed successfully on yeast P. pastoris and ChIFN-α was functional to stimulate anti- IBDV and NDV activities of cells.


Keywords


Pichia pastoris, chicken interferon-α, chicken interferon-γ

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DOI: https://doi.org/10.15625/0866-7160/v36n1se.4398

Published by Vietnam Academy of Science and Technology