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KMID : 0545120030130040537
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Journal of Microbiology and Biotechnology
2003 Volume.13 No. 4 p.537 ~ p.543
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Production of a Functional Mouse Interferon γ from Recombinant Saccharomyces cerevisiae
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LIM, YOUNG-YI
PARK, SEUNG-MOON/JANG, YONG-SUK/YANG, MOON-SIK/KIM, DAE HYUK
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Abstract
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Abstract The mouse interferon gcnc (MulFN-γ) was cloned and then used to transform Saccharonyces cerevisiae. Expressed MulFN-γ protein (MulFN-γ) was succcssfully secreted into culture medium due to the prcscnce of the signal peptide of rice amylasc IA. Two different promoters fused to MulFN-γ were tested: glyceraldehyde-3-phosphate dehydsogenase (GPD) promoter and a yeast hybrid ADHZ-GPD (AG) promoter consisting of alcohol dehydrogenase 11 (ADHZ) and GPD promoter. Using the hybrid promoter, the accumulation of MiiIFN- ytsanscript was the highest after 24 h cultivation, and then pdiially decreased the cultivation proceeded. However, both cell growth and recombinant MuiFN-γ production reached their peaks after the 4-day cultivation. It was possible to produce 6.5 mg/l of MuIFN-y without any changes in cell growth. Using GPD proinciter, the MulFN-γ transcript accumulation and the recombinant MuIFN-γ production followed the same pattem as the cell growth. However. compared Lo that of thc hybrid promoter, the priiduction of recombinant MuIFN-y was 0.2 mgL The secreted MulFN-y had estimaled molecular masses of 2I kDa and 13 kDa. which were larger than that of the encoded size due to glycosylation. The protection assay against the viral infection indicated that the recombinant MulFN-γ was bioactive.
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KEYWORD
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