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[摘要]
本研究采用毕赤酵母偏爱密码子,人工合成了高甜度Monellin基因,并构建了重组分泌型酵母表达载体pPIC9M,通过电击转化获得了可高效分泌表达有甜味活性高甜度Monellin的重组毕赤酵母GS115/pPIC9M。通过PCR检测证实Monellin基因已经整合进酵母基因组,SDS-PAGE和Western blot免疫杂交知所表达的蛋白是目的蛋白,最后通过双盲测定证实表达的蛋白存在正常活性。
[Key word]
[Abstract]
This experiment Pichia codon preference, a high sweetness synthetic gene Monellin, and the reorganization of production of yeast expression vector pPIC9M by electroporation can be a highly efficient expression of secretory activity sweet high sweetness Monellin reorganization Pichia GS115/pPIC9M. Monellin confirmed by PCR gene has been integrated into the yeast genome, SDS-PAGE and Western blot hybridization know immune expressed protein is the target protein final adoption of double-blind confirmed the presence of normal expression of activity.
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