Expression, purification, and bioactivity of a soluble recombinant ovine interferon-tau in Escherichia coli
Published Online: Jan 29, 2021
Page range: 101 - 108
Received: Jun 15, 2020
Accepted: Jan 27, 2021
DOI: https://doi.org/10.2478/jvetres-2021-0011
Keywords
© 2021 H.Y. Yu et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.
Introduction
Ovine interferon-tau (oIFN-τ) is a newly discovered type I interferon. This study used biochemical techniques to transform the oIFN-τ gene into
Material and Methods
First, total RNA was extracted from fresh sheep embryonic tissues with TRIzol reagent and then used as a template to reverse transcribe and amplify the mature oIFN-τ gene with RT-PCR. The amplified product was next digested with the
Results
The protein was confirmed to be oIFN-τ, which mainly existed in the soluble lysate fraction, as proven by SDS-PAGE and Western blot assays.
Conclusion
Purified IFN-τ exists mostly in a soluble form, and its anti-vesicular stomatitis virus (VSV) activity reached 7.08×10(6)IU/mL.